Full length human hcn1ih channel subunit and variants

ABSTRACT

This invention relates to a DNA sequence encoding a member of the hyperpolarised activated ion channel family (HCN) and variants thereof, and the use of said sequences in assays for the measurement of gene expression. It also relates to assays for screening of I h  activators and blockers for clinical and therapeutic use in the management of human psychiatric and neurological dysfunction in the CNS, cardiovascular dysfunction of the heart, and reproductive dysfunction and/or contraception related to I h  function in testes and spermatozoa. Further, antibodies against the expressed DNA sequences and other compounds reactive with the expressed DNA sequence are also part of the invention.

[0001] This invention relates to a DNA sequence encoding a member of thehyperpolarised activated ion channel family (HCN) and variants thereof,and the use of said sequences in assays for the measurement of geneexpression. It also relates to assays for screening of I_(h) activatorsand blockers for clinical and therapeutic use in the management of humanpsychiatric and neurological dysfunction in the CNS, cardiovasculardysfunction of the heart, and reproductive dysfunction and/orcontraception related to I_(h) function in testes and spermatozoa.Further, antibodies against the expressed DNA sequences and othercompounds reactive with the expressed DNA sequence are also part of theinvention

[0002] Voltage-gated ion channels play a critical role in shaping ofelectrical activity of neuronal and muscle cells, and in controlling thesecretion of neurotransmitters and hormones through the gating ofcalcium ion entry. Large families of voltage gated sodium (Na⁺),potassium (K⁺) and calcium (Ca²⁺) ion channels have been defined usingelectrophysiological, pharmacological and molecular techniques [1, 18];they are named according to their selective permeability for aparticular cation with reference to their voltage dependence, kineticbehaviour or molecular identity. The importance of membrane voltage andion permeability in the control of cell function ensures that modulationof ion channels will invariably have important consequences for cellsand tissues, and such modulation can often be turned to therapeuticadvantage. Major indication for ion channel modulators already includecardiac arrhythmia, hypertension, anxiety, epilepsy, pain,chemotherapy-induced nausea and diabetes as well as a range of drugs indevelopment for important new indications such as neuroprotection andpsychiatry.

[0003] A variety of bodily functions such as heart beat, sleep-wakecycles, secretion of hormones and control of behavioural state depend onthe action of pacemakers, specialised cells that are able to generaterhythmic, spontaneously firing action potentials. The archetypal organdisplaying autonomic rhythmicity is the heart.

[0004] Pacemaking in the heart is accomplished by the rhythmic dischargeof the sino atrial node [8, 11, 12]. The firing rate of the sino atrialnode is determined by the diastolic depolarisation phase of the actionpotential. During this phase the membrane potential is slowlydepolarised to the threshold triggering the next action potential. Theionic conductance underlying the cardiac pacemaker depolarisation wasidentified in the late seventies and early eighties [10] and calledI_(f) (f for ‘funny’) or I_(h) (h for hyperpolarisation activated). Asimilar current was subsequently discovered in neurones, first inphotoreceptors, [2, 3, 6] and then in various central neurons eghippocampal pyramidal cells [16] where is was called Iq (q for ‘queer’).This current was subsequently found in a wide variety of central andperipheral neurons [25].

[0005] I_(h) channels have several distinctive features. Unlike mostvoltage-gated channels, they open in response to negative-going voltagesteps to potentials within the range of the normal resting potential.They conduct both K⁺ and Na⁺ ions with a three fold greater permeabilityto K⁺, yielding a reversal potential of −30 to −40 mV underphysiological conditions. As a result, the opening of these channelsnear the resting potential (˜−60 mV) generates an inward, depolarisingcurrent that is largely carried by Na⁺[25]. Another unusual property ofthese channels is their regulation by cyclic nucleotides [11], whichspeed up the rate of channel activation by binding to an intracellularsite on the channel. In the heart, this in an important mechanismresponsible for the acceleration of heart rate in response tosympathetic stimulation. Activation of p-adrenergic receptors leads toan activation of adenylyl cyclase with the resulting increase inintracellular cAMP directly activating the I_(h) channel. cAMP bindingleads to a shift in the activation curve towards more positive voltages.This shift results in an increased inward current at a fixed membranepotential and therefore an acceleration of the diastolic depolarisation[9]. Muscarinic stimulation slows the heart rate, in part due to adecrease in cAMP level and a resulting reduction in the I_(h) current[13, 33].

[0006] In neurons, I_(h) channels have diverse functions. They wereinitially shown to be inward rectifiers; they are active near theresting potential and pass inward current more readily than outwardcurrent, thereby helping to control of resting potential and inputresistance [25]. In photoreceptors, I_(h) channels help to damp thehyperpolarising effect of light; they are activated byhyperpolarisation, causing the voltage response to light to fade duringthe first 100-200 ms, thus producing sensory adaptation. In many CNSneurons, activation of I_(h) channels following post inhibitory postsynaptic potentials contributes to a rebound afterdepolarisation (ADP),which can trigger an action potential. I_(h) is also generates orcontributes to ‘pacemaker’ potentials that controls the rate of rhythmicoscillations, similar to its role in the heart. I_(h) has been found toregulate the rhythmic activity of thalamic relay neurons [23] andinferior olivary neurons [5] through interaction with a T-type calciumcurrent. The oscillating single neurons are part of neuronal networksand are involved in the generation and modulation of rhythmic activityof these networks. A well studied example are spindle waves observed inthe EEG during slow wave sleep, which are generated through interactionsbetween thalamic reticular and relay neurons [4]. Regulation of I_(h) inthese cells is important in the sleep-wake cycle. Although less wellinvestigated, results suggest a similar role for I_(h) in the generationof oscillations in hippocampal neurons [21, 32] and respiratory neuronsof the preBotzinger complex of the ventrolateral medulla [26]. I_(h)channels are also expressed in dendrites where they influence the cableproperties of the dendrite and shape the time course of the EPSP as itis propagated to the soma [22]. A recent study has extended the role ofI_(h) neurons by showing that these channels can alter neurotransmitterrelease from presynaptic terminals as Crayfish neuromuscular synapses.Presynaptic cAMP generation by via serotonin receptor activationdirectly modulates I_(h) in axons that produces an increase in synapticstrength which cannot be explained solely by depolarisation of thepresynaptic membrane [7].

[0007] The genes encoding I_(h) channels were recently cloned from bothmammals [19, 28, 29] and sea urchins [15]. These genes, called HCN1-4 inmammals, are members of the voltage gated K⁺ channel family. The encodedproteins contain six transmembrane segments, including a positivelycharged S4 voltage sensor and a pore-forming P region that includes theK⁺ channel signature sequence GYG. In addition, the C-terminus containsa 120-amino acid sequence that is homologous to the cAMP and cGMPbinding domains of other proteins, and is therefore the likely site forcAMP regulation of channel opening [9,20]. All four mammalian genes areexpressed in brain, with differing expression patterns [24]. While thefirst reported cloning of I_(h) was from sea urchin spermatozoa [15],the functional significance is poorly understood at present. The channelis expressed in sperm flagellum and it was postulated that it may beinvolved in the control of flagella beating. One of the cloned isoforms,HCN4, has been detected in testes suggesting that I_(h) may also have afunction in mammalian testicular and sperm function [30].

[0008] These ion channels are the target for blocking molecules fortherapeutic use in dysfunction in the CNS, cardiovascular dysfunction ofthe heart, and reproductive dysfunction and/or contraception related toI_(h) function in testes and spermatozoa. For instance, compounds havealready been developed with the therapeutically interesting property ofinducing bradycardia with minimal inotropic side effects [14, 17].Unfortunately, there are no compounds yet described that distinguishcardiac from neuronal isoforms, and volunteers experienced opticalhallucinations probably due to reduced functionality of I_(h) inphotoreceptors. It is clear the ability to develop agents that areselective for the CNS vs heart or vice versa requires the availabilityof the cloned subunits to screen for compounds selective for subunitsexpressed in either neuronal or cardiac tissue.

[0009] It is the object of the present invention to provide full-lengthfunctional human HCN1 channel subunits. Furthermore, variants of thefull-length functional human HCN1 subunit are also provided. Human HCN1channel subunits are preferred over subunits from other species sincethey are preferably used to select compounds that can be used to treatCNS disorders, cardiovascular dysfunction of the heart, and reproductivedysfunction and/or contraception related to I_(h) function in testes andspermatozoa in humans.

[0010] One aspect of the invention concerns the full length cDNAsequence of the human HCN1 channel subunit (SEQ ID NO: 1). Theexpression of this subunit is predominantly restricted to the CNS andhas only been detected in some peripheral tissues to a very limitedextend. This subunit is therefore a candidate for screening compoundswith a selective effect in the CNS with a reduced propensity forundesirable side effects such as bradycardia via activity at Ih in theheart. In contrast with the prior art, the present invention providesthe full-length sequence of human HCN1 and sequence variants whereasprevious publications have only identified partial human HCN1 sequences.The major advantage of having a full-length sequence is that this allowsfunctional expression in in vitro systems (ie in transfected cell lines)for the identification of compounds increasing (opening the channel) ordecreasing (blocking the channel) ion flux through the expressedchannel. While expression of partial sequences may be possible, thesewill not give rise to channels with the full functional characteristicsof the full-length channel since they lack polypeptide domains thatcontribute to the functional characteristics of the channel. Inaddition, partial sequences may lack polypeptide domains with importanttarget sites for compounds that modulate channel activity. Functionalexpression of partial sequences, particularly those lacking the 5′endsuch as those reported for human HCN1 [1] is additionally problematicsince they lack the sequence encoding the N-terminal signal peptiderequired for directing the nascent polypeptide to the endoplasmicreticulum where all nascent plasma membrane bound proteins aresynthesised. The inherent advantage of an assay based on channelfunction, rather than, say a binding assay, is that a functional assayallows for the screening of compounds that interact with the channel aseither blockers or openers. Binding assays give no such information, nordo they allow the identification of allosteric or use-dependentmodulators of the channel.

[0011] By the term “full-length” as used herein we mean a DNA sequencethat contains a complete open reading frame, beginning with a startcodon at its 5′ end, preferably with a Kozak consensus, downstream to anin-frame stop codon.

[0012] The full-length DNA sequence for HCN1 (SEQ ID NO: 1) was obtainedusing a combined molecular biology and bioinformatics approach. Initialscreening of a proprietary database with the published human HCN1sequence [1] identified a single clone according to SEQ ID NO: 9 thatmatched at the 3′ end. Further analysis of the predicted amino acidsequence of this clone (SEQ ID NO: 10) confirmed that it encoded the 3′end of human HCN1 with an in-frame stop codon. Comparison with thefull-length murine HCN1 sequence indicated that the published humansequence lacks approximately 330 bases of 5′ sequence upstream to thetranslational initiation codon. Several approaches were adopted toattempt to obtain the missing 5′ end. These included cDNA libraryscreening, genomic library screening and 5′ RACE-PCR. RACE-PCRconsistently generated a further 153 bp (SEQ ID NO: 13) upstream of thepublished human HCN1 sequence. A search of genomic databases with thisnew 5′ end sequence identified a BAC clone (RP1 1-398G9) in the updatedEMBL high throughput genomic (HTG) database (em62htgnew). Sequenceanalysis of this clone suggests that it contains an in-frame initiationcodon and an open reading frame homologous to murine HCN1. The 5′ end ofhuman HCN1 is extremely GC-rich (approximately 80%) thus explaining whyconventional means of cloning this sequence proved fruitless.

[0013] Thus, sequence from a combination of newly identified genomic BACclone, RACE-PCR, published sequence and the clone obtained from theproprietary database have been assembled to generate a full length cDNAencoding a full length human HCN1 Ih channel subunit (SEQ ID NO: 1).

[0014] In another aspect of the invention, the sequences of the presentinvention can be used to derive primers and probes for use in DNAamplification reactions in order to perform diagnostic procedures or toidentify further, neighbouring genes which also may contribute to theexpression of HCN1. Also, fragments may be generated with PCRprocedures, as the sequences that are shown in SEQ ID NO: 13, SEQ ID NO:14, SEQ ID NO: 15, SEQ ID NO: 18, SEQ ID NO: 19 and SEQ ID NO: 21. Inthis way mutation analysis may be performed.

[0015] It is known in the art that genes may vary within and amongspecies with respect to their nucleotide sequence. The sequence of fulllength HCNI cDNAs from other individuals as well as their gene sequencemay now be readily identified using the above probes and primers.

[0016] Availability of the full-length sequence now allows to providefor assays that measure whether the functional characteristics of thefull-length channel are altered. It is clear for the skilled person thatnow all kinds of functional equivalents may be genetically engineered.Therefore, the invention also comprises functional equivalents, whichare characterised in that they are capable of hybridising to at leastpart of the HCNI sequence shown in SEQ ID NO: 1, preferably under highstringency conditions.

[0017] The present invention also includes these functional variants.Examples of such variants are for example deletions, insertions, pointsubstitutions (single nucleotide polymorphisms) and splice variants.Also included in this term are other sequence variants, for examplesplice variants of a sequence which may differ in size through inclusionor exclusion of different exons. Identification of sequence variants andsingle nucleotide polymorphisms in particular are important for tworeasons: first by identifying the small differences that influence thetype and severity of diseases contracted by individuals; and second, bybeing able to understand the reasons behind variation of response tomedicines at the individual level. Understanding how these variations inthe genetic code influence biological systems will be the key todiscovering new medicines and enabling doctors to prescribe medicines topatients who are likely to respond.

[0018] The present invention provides such functional variants. SEQ IDNO: 3 which is a full-length variant that contains a 189 bp insertion atposition 2114 in SEQ ID NO 1, indicative of a splicing variant comparedto SEQ ID NO: 1. This insertion encodes a further 63 amino acids in SEQID NO 4 in a region predicted to encode a portion of thecarboxy-terminal domain of the polypeptide (FIGS. 1 and 2). Two singlenucleotide variants are also provided in SEQ ID NO 5. The first is a C(SEQ ID NO 1) to A (SEQ ID NO 5) transition at position 107 resulting ina change of amino acid from Pro (SEQ lD NO 2) to Thr (SEQ ID NO 6) atposition 28. This occurs in a region predicted to encode the aminoterminal domain of the polypeptide (FIGS. 1 and 2). The second singlenucleotide variant is a C (SEQ ID NO 1) to T (SEQ ID NO 5) transition atposition 2064 resulting in a change of amino acid from Ser (SEQ ID NO 2)to Phe (SEQ ID NO 6). This occurs in a region predicted to encode thecarboxy-terminal domain of the polypeptide immediately upstream of thesplice variant insertion site. Both single nucleotide variants maytherefore occur in association with the splicing variant giving rise toSEQ ID NO 7 encoding the polypeptide SEQ ID NO 8. In addition, afunctional variant may comprise only one of these single nucleotidedifferences in conjunction with the splicing isoforms.

[0019] In addition, the sequences of the present invention contain anumber of other differences compared to the published sequence [1].These include a further eleven point substitutions, corresponding to 5amino acid changes, and divergent sequence at the 3′ end of thepublished sequence [1].

[0020] Two nucleic acid fragments are considered to have hybridisablesequences if they are capable to hybridising to one another undertypical hybridisation and wash conditions, as described, for example inManiatis, et al., pages 320-328, and 382-389, or using reducedstringency wash conditions that allow at most about 25-30% basepairmismatches, for example: 2×SSC, 0.1% SDS, room temperature twice, 30minutes each, then 2×SSC, 0.1% SDS 37° C. once, 30 minutes; then 2×SSC,room temperature twice ten minutes each. Preferably, homologous nucleicacid strands contain 15-25% basepair mismatches, even more preferably5-15% basepair mismatches. These degrees of homology can be selected byusing wash conditions of appropriate stringency for identification ofclones from gene libraries or other sources of genetic material, as iswell known in the art.

[0021] Furthermore, to accommodate codon variability, the invention alsoincludes sequences coding for the same amino acid sequences as thesequences disclosed herein. Also portions of the coding sequences codingfor individual domains of the expressed protein are part of theinvention as well as allelic and species variations thereof. Sometimes,a gene expresses different isoforms in a certain tissue which includessplicing variants, that may result in an altered 5′ or 3′ mRNA or in theinclusion of an additional exon sequence. Altematively, the messengermight have an exon less as compared to its counterpart. These sequencesas well as the proteins encoded by these sequences all are expected toperform the same or similar functions and form also part of theinvention.

[0022] The sequence information as provided herein should not be sonarrowly construed as to require inclusion of erroneously identifiedbases. The specific sequence disclosed herein can be used to isolatefurther genes which in turn can be subjected to further sequenceanalyses thereby identifying sequencing errors.

[0023] Thus, in one aspect, the present invention provides for a DNAsequence encoding a full length human hyperpolarised activated ionchannel of the HCN 1 subtype or functional equivalents thereof.

[0024] The DNA according to the invention may be obtained from cDNA.Alternatively, the coding sequence might be obtained from genomic DNA,or prepared using DNA synthesis techniques. The polynucleotide may alsobe in the form of RNA. The polynucleotide may be in single stranded ordouble stranded forn. The single strand might be the coding strand orthe non-coding (anti-sense) strand.

[0025] The present invention further relates to polynucleotides whichhave at least 80%, preferably 90% and more preferably 95% and even morepreferably at least 98% identity with SEQ ID NO:1, provided that suchpolynucleotides encode polypeptides which retain essentially the samebiological function or activity as the natural, mature protein. Evenmore preferred is the full length sequence according to SEQ ID NO: 1,SEQ ID NO: 3, SEQ ID NO 5 or SEQ ID NO 7

[0026] The percentage of identity between two sequences can bedetermined with programs such as DNAMAN (Lynnon Biosoft, version 3.2).Using this program two sequences can be aligned using the optimalalignment algorithm of Smith and Waterman (1981, J. Mol. Biol,147:195-197). After alignment of the two sequences the percentageidentity can be calculated by dividing the number of identicalnucleotides between the two sequences by the length of the alignedsequences minus the length of all gaps.

[0027] The DNA according to the invention will be very useful for invivo or in vitro expression of the novel sequence according to theinvention in sufficient quantities and in substantially pure form.

[0028] In another aspect of the invention, a full length humanhyperpolarised activated ion channel of the HCN 1 subtype or functionalequivalents thereof are provided. An example of such a polypeptide is apolypeptide sequence according to SEQ ID NO: 2. SEQ ID NO: 2 is encodedby the open reading frame of SEQ ID NO: I and consists of 827 aminoacids.

[0029] Preferably, the polypeptides according to the invention comprisethe amino acid sequence as shown in SEQ ID NO: 2.

[0030] Also functional equivalents, that is polypeptides homologous toSEQ ID NO: 2 or parts thereof having variations of the sequence whilestill maintaining functional characteristics, are included in theinvention. Such variants are provided in SEQ ID NO: 4 which is apolypeptide encoded by the open reading frame of SEQ ID NO: 3, SEQ IDNO: 6 which is a polypeptide encoded by the open reading frame of SEQ IDNO: 5 and SEQ ID NO: 8 which is a polypeptide encoded by the openreading frame of SEQ ID NO: 7

[0031] The variations that can occur in a sequence may be demonstratedby (an) amino acid difference(s) in the overall sequence or bydeletions, substitutions, insertions, inversions or additions of (an)amino acid(s) in said sequence. Amino acid substitutions that areexpected not to essentially alter biological and immunologicalactivities, have been described. Amino acid replacements between relatedamino acids or replacements which have occurred frequently in evolutionare, inter alia Ser/Ala, Ser/Gly, Asp/Gly, Asp/Asn, IleNal (see Dayhof,M. D., Atlas of protein sequence and structure, Nat. Biomed. Res.Found., Washington D.C., 1978, vol. 5, suppl. 3). Based on thisinformation Lipman and Pearson developed a method for rapid andsensitive protein comparison (Science, 1985, 227, 1435-1441) anddetermining the functional similarity between homologous polypeptides.It will be clear that also polynucleotides coding for such variants arepart of the invention.

[0032] The polypeptides according to the present invention include thepolypeptides comprising SEQ ID NO: 2 but also its isoforms, i.e.polypeptides with a similarity of 70%, preferably 85%, more preferably90% even more preferably 95% or even 98%, provided these isoforms retainthe same biological functions as the sequence shown in SEQ ID NO: 2,like the sequences shown in SEQ ID NO: 4, SEQ ID NO: 6 and SEQ ID NO: 8.Also fragments of such polypeptides still capable of conferring thesebiological effects are included. Especially portions which still bind toligands form part of the invention. Such portions may be functional perse, e.g. in solubilized form or they might be linked to otherpolypeptides, either by known biotechnological ways or by chemicalsynthesis, to obtain chimeric proteins. Such proteins might be useful astherapeutic agent in that they may substitute the gene product inindividuals with aberrant expression of the HCN1 gene. Polypeptidesobtainable from alternative splice products are also included in thisinvention.

[0033] As an example of a functional equivalent SEQ ID NO: 4 isprovided. This polypeptide corresponds to the open reading frame of SEQID NO: 3 and contains an insertion of 63 amino acids.

[0034] Further examples of functional variants are polypeptides derivedfrom coding sequences with single nucleotide variations. These includeSEQ ID NO: 6 and SEQ ID NO: 8 which correspond to the open readingframes of SEQ ID NO: 5, and SEQ ID NO: 7 respectively. Both polypeptidesincorporate amino acid substitutions at position 28 (Pro/Thr) and 680(Ser/Phe). It should be noted that either amino acid substitution mayoccur alone in the HCN1 polypeptide sequence such that any HCN1polypeptide with either one or both of these substitutions are part ofthe invention.

[0035] The sequence according to SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO:5 or SEQ ID NO: 7 may also be used in the preparation of vectormolecules for the expression of the encoded protein in suitable hostcells. Also, fragments of these sequences may be cloned into anexpression vector and expressed in order to determine individualfunctional sites along the cDNA molecule. The sequences may also bemodified in that they are cloned into an expression vector either withour without the 5′ and/or 3′ non-coding regions. The coding region of acDNA clone according to SEQ ID NO: 1 is shown in SEQ ID NO: 21. Thecoding region of a cDNA clone according to SEQ ID NO: 3 is shown in SEQID NO: 22. The coding region of a cDNA clone according to SEQ ID NO: 5is shown in SEQ ID NO: 24. The coding region of a cDNA clone accordingto SEQ ID NO: 7 is shown in SEQ ID NO:25. The invention therefore alsoprovides an expression vector comprising SEQ ID NO: 22, SEQ ID NO: 23,SEQ ID NO: 24 or SEQ ID NO: 25 or fragments thereof.

[0036] A wide variety of host cell and cloning vehicle combinations maybe usefully employed in cloning the nucleic acid sequence coding for theHCN1gene, the cDNA coding for HCN1 or parts thereof. For example, usefulcloning vehicles may include chromosomal, non-chromosomal and syntheticDNA sequences such as various known bacterial plasmids and wider hostrange plasmids and vectors derived from combinations of plasmids andphage or virus DNA.

[0037] Vehicles for use in expression of the genes or a ligand-bindingdomain thereof of the present invention will further comprise controlsequences operably linked to the nucleic acid sequence coding for aligand-binding domain. Such control sequences generally comprise apromoter sequence and sequences which regulate and/or enhance expressionlevels. Of course control and other sequences can vary depending on thehost cell selected.

[0038] Suitable expression vectors are for example bacterial or yeastplasmids, wide host range plasmids and vectors derived from combinationsof plasmid and phage or virus DNA. Vectors derived from chromosomal DNAare also included. Furthermore an origin of replication and/or adominant selection marker can be present in the vector according to theinvention. The vectors according to the invention are suitable fortransforming a host cell.

[0039] Recombinant expression vectors comprising the DNA of theinvention as well as cells transformed with said DNA or said expressionvector also form part of the present invention.

[0040] Suitable host cells according to the invention are bacterial hostcells, yeast and other fungi, plant or animal host such as ChineseHamster Ovary cells or monkey cells. Especially preferred cells are HEK293. Thus, a host cell which comprises the DNA or expression vectoraccording to the invention is also within the scope of the invention.The engineered host cells can be cultured in conventional nutrient mediawhich can be modified e.g. for appropriate selection, amplification orinduction of transcription. The culture conditions such as temperature,pH, nutrients etc. are well known to those ordinary skilled in the art.

[0041] The techniques for the preparation of the DNA or the vectoraccording to the invention as well as the transformation or transfectionof a host cell with said DNA or vector are standard and well known inthe art, see for instance Sambrook et al., Molecular Cloning: Alaboratory Manual. 2^(nd) Ed., Cold Spring Harbor Laboratory, ColdSpring Harbor, N.Y., 1989.

[0042] The proteins according to the invention can be recovered andpurified from recombinant cell cultures by common biochemicalpurification methods including ammonium sulfate precipitation,extraction, chromatography such as hydrophobic interactionchromatography, cation or anion exchange chromatography or affinitychromatography and high performance liquid chromatography. If necessary,also protein refolding steps can be included.

[0043] The gene products according to the present invention can be usedfor the in vivo or in vitro identification of novel ligands or analogsthereof. For this purpose binding studies can be performed with cellstransformed with DNA according to the invention or an expression vectorcomprising DNA according to the invention, said cells expressing theHCN1 full length gene product according to the invention or fragmentsthereof.

[0044] Alternatively also the HCN1 gene products according to theinvention as well as ligand-binding domains thereof can be used in anassay for the identification of functional ligands or analogs for theHCN1 gene products.

[0045] Methods to determine binding to expressed gene products as wellas in vitro and in vivo assays to determine biological activity of geneproducts are well known. In general, expressed gene product is contactedwith the compound to be tested and binding, stimulation or inhibition ofa functional response is measured.

[0046] As a preferred way of detecting the binding of the ligand to theexpressed protein, also signal transduction capacity may be measured.

[0047] The present invention thus provides for a quick and economicmethod to screen for therapeutic agents for the prevention and/ortreatment of diseases related to CNS disorders such as human psychiatricand neurological dysfunction, cardiovascular dysfunction of the heart,and reproductive dysfunction and/or contraception related to I_(h)function in testes and spermatozoa. The method is especially suited tobe used for the high throughput screening of numerous potentialcompounds.

[0048] Compounds which activate or inhibit the function of HCN1 geneproduct may be employed in therapeutic treatments to activate or inhibitthe polypeptides of the present invention.

[0049] Also within the scope of the invention are antibodies, especiallymonoclonal antibodies raised against the polypeptide molecule accordingto the invention. Such antibodies can be used therapeutically to inhibitHCN1 gene product function and diagnostically to detect HCN1 geneproducts.

[0050] The invention furthermore relates to the use of the HCN1 geneproducts as part of a diagnostic assay for detecting clinicalabnormalities or susceptibility to any of the above disorders orinvestigation of different clinical outcomes in response to medicaltreatment related to mutations in the nucleic acid sequences encodingthe HCN1 gene. Two examples single nucleotide variants are provided inthis invention. Such variants or mutations may e.g. be detected by usingPCR (Saiki et al., 1986, Nature, 324, 163-166). Also the relative levelsof RNA can be determined using e.g. hybridization or quantitative PCRtechnology. The presence and the levels of the HCN1 gene productsthemselves can be assayed by immunological technologies such asradioimmuno assays, Western blots and ELISA using specific antibodiesraised against the gene products. Such techniques for measuring RNA andprotein levels are well known to the skilled artisan.

[0051] The determination of expression levels of the HCNI gene productsand variants thereof in individual patients may lead to fine tuning oftreatment protocols.

[0052] Also, transgenic animals may be prepared in which the expressionof the HCN1 gene is altered or abolished.

EXAMPLE 1

[0053] Full-length Sequence identification

[0054] Proprietary databases were screened by BLAST2 for the presence ofrelated human cDNA sequences using parts of the DNA sequence of thehuman HCNI channel (Accession No AF064876) [29] and mouse HCN1(Accession number AJ225123). A single cDNA clone (SEQ ID NO: 9) wasidentified from a brain cDNA library, obtained and sequenced using anABI Prism 310 Genetic analyser (PE Biosystems). Sequencing reactionswere performed using ABI Prism BigDye Terminator cycle sequencing Readyreaction kit (PE Biosystems). Each sequencing reaction contained 300ngcDNA clone, 3.2pmol sequencing primer, and PE Biosystems TerminatorReady reaction mix in a final volume of 20ul. Reactions were cycled asfollows: 25 cycles of 96° C. for 10 sec, 50° C. for 5sec and 60° C. for4min in a PE Biosystems GeneAmp PCR system 9700. Following cycling, theextension products were precipitated by adding 2 ul 3M NaOAc (pH 4.6)and 50 ul 95% ethanol. Products were precipitated at RT for 15 min andcollected by centrifugation at 14000 rpm for 20 min. Pellets were washed2× with 70% ethanol prior to resuspension in 20 ul

[0055] The published human HCN1 sequence and the 5′ sequence obtained byRACE-PCR was used to design primers to PCR the region of human HCNIflanked by the RACE-PCR product (SEQ ID NO: 13) and SEQ ID NO: 9 andcovered by the published sequence. Each PCR reaction contained 1×PCRbuffer (Expand High Fidelity buffer), 1.5 mM MgCl₂, 2 μl whole brainMarathon-Ready cDNA (Clontech), 400 nM primer 1 (HCNIPCRA5′-TGCTGCAGCCCGGGGTCAACMAT-3′), 400 nM primer 2 (HCN1PCRB5′-GAGGCGGTGGGGGAGGCATAGTGG-3′), 400 μM dATP, 400 μM dCTP, 400μM dGTP,400 μM dTTP, 5% DMSO and 2.625 units Expand High Fidelity PCR enzyme mixin a total volume of 50 μl. Reactions were cycled in a MJ ResearchPTC-200 Thermal Cycler using the following conditions: 94° C., 2 min and35 cycles of 24° C. for 30sec, 60° C. for 30 sec, 72° C. for 90 sec,followed by an extension of 72° C. for 5 min. A PCR product ofapproximately 1.9 kb was identified, purified and sequenced aspreviously described (SEQ ID NO: 19). Some independently derived cloneswere also found to contain a single nucleotide variation (a C to Ttransition) indicated in SEQ ID NO: 20

[0056] The full length sequence of HCN1 indicates that the cDNA consistsof 2484bp open reading frame (SEQ ID NO: 22) encoding an 827 amino acidchannel. The functional equivalent sequence shown in SEQ ID NO: 3comprises an 2673bp open reading frame (SEQ ID NO: 23) encoding a 890amino acid channel. Two single nucleotide variants were found whichaltered the amino acid sequence, and these are indicated in conjunctionwith the splice variant in SEQ ID NO: 24 and SEQ ID NO: 25. To date,four members of the hyperpolarisation-activated cation channel have beenidentified and cloned from mouse (HCN14). Full length clones of humanHCN-2 and HCN4 (Accession NOs AF065164 and AJ238850) have also beenobtained whereas only a single partial sequence exists for human HCN-1.These channels share close overall homology. Analysis of the channelsequence have shown that these channels have the classic K⁺ channelstructure of 6 transmembrane domains and an ion conducting pore loop.Additionally, they possess a cyclic nucleotide binding domain. Withinthe transmembrane, pore and CNBD regions there is very high conservationamong the channel subtypes. Alignment of the HCN1 sequences according tothe invention with the other members of the HCN channel family suggestthat this sequence does encode an hyperpolarisation activated ionchannel. Regions of conservation between the human HCN channels aredenoted by asterixes in FIG. 1. The transmembrane domains, pore regionsand CNBD are underlined. Alignment of HCNI polypeptides shows that thenovel sequences do encode HCN1 channels (FIG. 2). The single amino acidvariants are indicated in bold and are underscored with a # in thealignment in FIG. 2.

[0057] The cloned full length mouse HCNI channel contains apolyglutamine repeat in the C-terminal region [291. Expansions ofpolyglutamine repeats have been implicated in several neurodegenerativediseases [27]. Although no polyglutamine repeat appears to be present inthe novel sequence the possibility arises of allelic variation andexpansion of a polyglutamine tract resulting in increased propensity toneurodegenerative diseases. The 189 bp deletion identified in the newsplice variant spans the corresponding region in the mouse HCNI channelcontaining the polyglutamine repeat.

EXAMPLE 2

[0058] Tissue distribution of SEQ ID NO: 1 and SEQ ID NO: 3

[0059] In order to analyze the expression of the ion channels comprisingSEQ ID NO:1 and SEQ ID NO: 3 in different human tissues, Northern blotanalysis was performed. A human Multiple Tissue Expression (MTE) Arraywas obtained from Clontech, which contains poly A⁺ RNA from a variety ofhuman tissues including nervous system, cardiovascular, digestive, andimmune tissue. Prehybridization was performed in Clontech's ExpressHybsupplemented with 100 μg/ml denatured sheared salmon DNA for 2 hr at 65°C. For RNA detection, ³²P labeled DNA fragments were generated with anOligolabeling kit (High Prime, Boehringer Mannheim) using a 627 bpfragment generated from SEQ ID NO: 9. This fragment starts at a Smalsite 717 bp from the 5′ end of the coding sequence and stops at the Notlcloning site (SEQ ID NO:21) and is common to both SEQ ID NO: 1 and SEQID NO: 3. The ³²P labeled probes were hybridized to the array for 16hours at 65° C. Subsequently, the array was washed with 2×SSC/1% SDS at65° C. (4×20 min) followed by 2×20 min washes at 50° C. in 0.1×SSC/0.5%SDS. Filters were exposed to a phosphorscreen for 20 hr and analysedusing Molecular Dynamics Storm Scan phosphorimage package.

[0060] From the results it can be concluded that human SEQ ID NO: 1 andSEQ ID NO: 3 are expressed predominantly in nervous tissue. Positivesignals were detected in a variety of brain regions including cerebralcortex, cerebellum, occipital lobe, temporal lobe and nucleus accumbens,Weaker expression was observed in fetal brain. Peripherally, only weakexpression was detected in atrial tissue of the heart.

EXAMPLE 3

[0061] Tissue distribution of SEQ ID: 1 and 3 in nervous tissue

[0062] In order to further analyze the expression of the ion channelscomprising SEQ ID NO:1 and SEQ ID NO: 3 in nervous tissue, Northern blotanalysis was performed. A human Multiple Tissue Northern was obtainedfrom Clontech (Human Brain MTN Blot II), which contains poly A⁺ RNA froma variety of brain regions. Prehybridization was performed in Clontech'sExpressHyb for 2 hr at 65 ° C. For RNA detection, ³²P labeled DNAfragments were generated as described previously for MTE array. The ³²Plabeled probes were hybridized to the filters for 16 hours at 65° C.Subsequently, the filters were washed with 2×SSC/0.05% SDS at roomtemperature (2×40 min) followed by 2×20 min washes at 50° C. in0.1×SSC/0.1% SDS. Filters were exposed to a phosphorimager screen for20hr and analysed using Molecular Dynamics Storm Scan phosphorimagepackage.

[0063] From the results it can be concluded that human SEQ ID NO: 1 andSEQ ID NO: 3 are expressed in a variety of nervous tissue. Multipletranscripts were detected in a cerebral cortex, cerebellum, occipitallobe, temporal lobe and frontal lobe. Weaker expression was observed inputamen and medulla, with no expression in spinal cord.

EXAMPLE 4

[0064] Functional assay for detecting HCNI channel currents

[0065] Human HCN1 channel currents were detected under voltage-clampusing the whole-cell configuration of the patch-clamp technique.Appropriate cells (HEK293) expressing human HCN1 channel subunits wereplaced in a recording chamber containing extracellular solution (Sodiumchloride 135 mM; Potassium chloride 5 mM; Calcium chloride 1.8 mM;Magnesium chloride 0.5 mM; HEPES 5 mM) at a temperature of 20-25 degreescentigrade. In some experiments the extracellular solution contained 30mM Potassium chloride, in which cases the Sodium chloride concentrationwas reduced to 110 mM. The glass pipettes used for recording containedpipette solution (Sodium chloride 10 mM; Potassium chloride 130 mM;Magnesium chloride 0.5 mM; HEPES 5 mM; EGTA 1 mM) Using standardequipment and methodology, a cell was voltage-clamped at a holdingpotential of 40 mV (not allowing for liquid junction potentials). Themembrane current was recorded throughout the experiment. To determinethe current-voltage relationship of human HCN1 channels, every 8 secondsthe membrane potential was stepped sequentially to the following values(in mV) for 2 or 3 seconds each: −20, 40, −60, −80, −100, −120, −140(FIGS. 5 and 6). To determine the effects of caesium chloride (5 mM) orZD 7288 (100 micromolar) on human HCN1 channel currents, t every 8 or 10seconds the cell potential was hyperpolarised to a potential of −120 mV(not allowing for liquid junction potentials) for 1-5 seconds and thepotential then returned to the holding potential of −40 mV. The membranecurrent was measured just after (1-10 milliseconds) the start of thehyperpolarising step and just before (10-100 milliseconds) its end. Thedifference in these two values of membrane current was taken as theamplitude of the current due to the human HCN1 channel subunits. Foreach cell, the current amplitude was determined first in normalextracellular solution and then in extracellular solution containingcaesium chloride (FIG. 3) or ZD 7288 (FIGS. 4 and 7). Alternatively, theeffect of caesium chloride (5 mM) on human HCN1 channel currents wasdetermined by measuring the current-voltage relationship in normalextracellular solution and also in extracellular solution containingcaesium chloride (FIG. 6). Human HCN1 channel currents were reduced bycaesium chloride (5 mM) and by ZD 7288 (100 micromolar) [17, 25].

[0066] A clone which has the following characteristics is considered tobe an I_(h) ion channel:

[0067] Template suppression reagent (PE Biosystems) for sequencing. Thisclone encoded the 3′ end of human HCN-1. The sequence is shown in SEQ IDNOs: 9.

[0068] The presence of a second splice variant was confirmed by PCRusing primers designed against the published sequence flanking eitherside of the 189bp deletion identified in SEQ ID NO: 9 compared to thepublished sequence. Each PCR reaction contained 1×PCR buffer (ExpandHigh Fidelity buffer), 1.5 mM MgCl₂, 2 μl whole brain Marathon-ReadycDNA (Clontech), 400 nM primer 1 and 2 (HCN1SPLICEA5′-TGCTGCAGCCCGGGGTCAACAAAT-3′ and HCNISPLICE B5′-CTCCTGCCCCCTGCCTGAAG-3′, or HCN1SPLICEC 5′-TCTACTACGACCCCGACCTC-3′and HCN1 SPLICED 5′-TGGCTCCCGACGACATCT-3′), 400 μM dATP, 400 μM dCTP,400 μM dGTP, 400 μM dTTP, 5% DMSO and 2.625 units Expand High FidelityPCR enzyme mix in a total volume of 50 μl. Reactions were cycled in a MJResearch PTC-200 Thermal Cycler using the following conditions: 94° C.,2min and 35 cycles of 24° C. for 30sec, 60° C. for 30 sec, 72° C. for 90sec, followed by an extension of 72° C. for 5 min. PCR products ofapproximately 560 bp (HCNISPLICEA and HCN1SPLICEB) and 600 bp(HCN1SPLICEC and HCN1SPLICED) were identified, purified and sequenced aspreviously described (SEQ ID NO: 17). This sequence had no deletioncompared to the published sequence. Several independently derived cloneswere observed to contain a single nucleotide variant (a C to Ttransition) shown in SEQ ID NO: 11 which results in a change in aminoacid (Ser to Phe) in SEQ ID NO: 12.

[0069] Primers were designed using the known human HCN1 sequence toattempt to obtain the 5′ end of HCN1 by RACE-PCR using Clontech SMARTRACE cDNA amplification kit. SMART 5′-RACE-ready cDNA was synthesised asfollows, 1 μg of hippocampal polyA⁺ RNA was mixed with 2 μM 5′-RACE cDNAsynthesis primer (Clontech 5′-(T)₂₅N⁻¹,N-3′) and 2uM SMART IIoligonucleotide (Clontech 5′-AAGCAGTGGTMCAACGCAGAGTACGCGGG-3′) in atotal volume of 5 μl. Tubes were incubated at 70° C. for 2 min andcooled on ice for a further 2min. The following was then added, 1×Firststrand buffer (50 mM Tris-HCl pH8.3, 75mM KCl and 6 mM MgCl₂), 2 mM DTT,1 mM dATP, 1 mM dCTP, 1 mM dGTP, 1 mM dTTP and 200 u MMLV reversetranscriptase. The reactions were incubated at 42° C. for 1.5hr in anair incubator. The reaction product was diluted with 250 ul Tricine-EDTAbuffer and heated at 72° C. for 7 min prior to storage at −20° C.

[0070] Each RACE-PCR reaction contained 1 x PCR buffer (ClontechAdvantage-GC 2 PCR buffer, 40 mM Tricine-KOH pH9.2, 15 mM KOAc, 3.5 mMMg(OAc)₂, 5% DMSO, 187.5 ng BSA, 0.005% Nonidet P-40 and0.005%Tween-20), 200 uM dATP, 200 uM dCTP, 200 uM dGTP, 200 uM dTTP, 2.5ul RACE-ready cDNA, Clontech Universal primer mix (2OnM long primer5′-CTMTACGACTCACTATAGGGCAAGCAGTGGTMCAACGCAGAGT-3′, 100 nM short primer5′-AAGCAGTGGTMCMCGCAGACT-3′), 200 nM Gene Specific Primer (BCNG1 R2, 5′-CTGGCTGTCTTGTAAACTTCAGATCCATT-3′, BCNG1R35′-CTGTMGGGTGGATMTCCAGAAGCCTGC-3′, BCNG R4B,5′-TTCTGGCTCCCAAACATGCGGAGG-3′), 1×Advantage-GC 2 polymerase mix (1%glycerol, 0.3 mM Tris-HCl pH 8.0, 1.5 mM KCl, 1 uM EDTA) and 0.5MGC-melt (Clontech) in a total volume of 50 ul. Reactions were cycled ina MJ Research PTC-200 Thermal Cycler using the following conditions: 95°C., 3 min and 40 cycles of 94° C. for 20 sec, 65° C. for 20 sec, 72° C.for 90sec, followed by an extension of 72° C. for 5min. A PCR product ofapproximately 300 bp was identified, purified and sequenced aspreviously described. RACE-PCR generated novel sequence information of153bp upstream of published human HCN1 sequence SEQ ID NO: 13.

[0071] SEQ ID NO: 13 was used to screen by BLAST2 human public EST andgenomic databases available from EMBL [31]. A human genomic bacterialartificial chromosome (BAC) clone (RP11 -398G9: accession numberAC013384) that contained sequence similarity was identified from thehigh throughput genomic division of the EMBL release 62. The regioncorresponding to human HCN-1 was sequenced as described previously. Thisclone appeared to encode the 5′ end of human HCN-1 including thetranslation initiation codon. The sequences are shown in SEQ ID NO: 14.Several independently derived clones were found to have a singlenucleotide variant (a C to A transition) indicated in SEQ ID NO: 15resulting in a change of amino acid from Pro to Thr.

[0072] 1. It opens through hyperpolarisation and closes at positivevoltage values (V_(m)-10 mV;

[0073] 2. Whose activation and deactivation proceeds with a relativelyslow time course;

[0074] 3. Which conducts not only K⁺ ions but also Na⁺ ions;

[0075] 4. Which are blocked preferentially by extracellular caesium ionsrather than by extracellular barium ions.

[0076] 5. Which are directly modulated by cyclic nucleotides,particularly cAMP and cGMP

LEGENDS TO THE FIGURES

[0077]FIG. 1: CLUSTAL W Multiple Sequence Alignments of SEQ ID NO: 2 SEQID NO: 4, SEQ ID NO: 6 and SEQ ID NO: 8 with other human HCN channels

[0078]FIG. 2: CLUSTAL W Multiple Sequence Alignments of SEQ ID NO: 2,SEQ ID NO: 4, SEQ ID NO: 6 and SEQ ID NO: 8 with rat, mouse and partialhuman HCN1 channels

[0079]FIG. 3: Effect of caesium chloride (5 mM) on hHCN1 (full-length)currents. Whole- cell patch-clamp recording from a HEK 293F celltransfected with an expression vector containing SEQ ID NO 7. Panel A:each data point represents a measurement from one data sweep. Caesiumchloride was added to the bathing medium for the time represented by thebar above the data points. Panel B: representative data sweeps(corresponding letters in panels A & B identify the times at which thesweeps in panel B were taken). Inward membrane currents in panel B areplotted as negative quantities (following convention), but the amplitudeof hHCN1 current in panel A is plotted as a positive value.

[0080]FIG. 4: Effect of ZD 7288 (100 micromolar) on hHCN1 (full-length)currents. Whole-cell patch-clamp recording from a HEK 293F celltransfected with an expression vector containing SEQ ID NO 7. Panel A:each data point represents a measurement from one data sweep. ZD 7288was added to the bathing medium for the time represented by the barabove the data points. Panel B: representative data sweeps(corresponding letters in panels A & B identify the times at which thesweeps in panel B were taken). Inward membrane currents in panel B areplotted as negative quantities (following convention), but the amplitudeof hHCN1 current in panel A is plotted as a positive value.

[0081]FIG. 5: Current-voltage relationships of whole-cell patch-clamprecordings from HEK 293F cells transfected with DNA for hHCN1. Panel A:SEQ ID NO 5. Panel B:

[0082] SEQ ID NO 7. For each panel, the upper traces display thecurrents evoked by the voltage steps shown in the lower traces.

[0083]FIG. 6: Current-voltage relationship and block by caesium chlorideof hHCN1 current. Whole-cell patch-clamp recording from a HEK 293F celltransfected with DNA expressing SEQ ID NO:3. Each panel shows thecurrents evoked by voltage commands (voltage traces not shown). Panel A:normal extracellular solution. Panel B: extracellular solutioncontaining 5 mM caesium chloride. Panel C: return to normalextracellular solution. All panels from the same cell.

[0084]FIG. 7: Effect of ZD 7288 (100 micromolar) on hHCN1 current.Whole-cell patch-clamp recording from a HEK 293 cell transfected witKDNA expressing SEQ ID NO:3. Panel A: each data point represents ameasurement from one data sweep. ZD 7288 was added to the bathing mediumfor the time represented by the bar above the data points. Panel B:representative data sweeps (corresponding letters in panels A & Bidentify the times at which the sweeps in panel B were taken). Note thatinward membrane currents in panel B are plotted as negative quantities(following convention), but that the amplitude of hHCN1 current in panelA is plotted as a positive value.

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1 25 1 2791 DNA Homo sapiens 1 ccgtcgccgg ccgcgtcctc cgggcatggaaggaggcggc aagcccaact cttcgtctaa 60 cagccgggac gatggcaaca gcgtcttccccgccaaggcg tccgcgccgg gcgcggggcc 120 ggccgcggcc gagaagcgcc tgggcaccccgccggggggc ggcggggccg gcgcgaagga 180 gcacggcaac tccgtgtgct tcaaggtggacggcggtggc ggcggtggcg gcggcggcgg 240 cggcggcgag gagccggcgg ggggcttcgaagacgccgag gggccccggc ggcagtacgg 300 cttcatgcag aggcagttca cctccatgctgcagcccggg gtcaacaaat tctccctccg 360 catgtttggg agccagaagg cggtggaaaaggagcaggaa agggttaaaa ctgcaggctt 420 ctggattatc cacccttaca gtgatttcaggttttactgg gatttaataa tgcttataat 480 gatggttgga aatctagtca tcataccagttggaatcaca ttctttacag agcaaacaac 540 aacaccatgg attattttca atgtggcatcagatacagtt ttcctattgg acctgatcat 600 gaattttagg actgggactg tcaatgaagacagttctgaa atcatcctgg accccaaagt 660 gatcaagatg aattatttaa aaagctggtttgtggttgac ttcatctcat ccatcccagt 720 ggattatatc tttcttattg tagaaaaaggaatggattct gaagtttaca agacagccag 780 ggcacttcgc attgtgaggt ttacaaaaattctcagtctc ttgcgtttat tacgactttc 840 aaggttaatt agatacatac atcaatgggaagagatattc cacatgacat atgatctcgc 900 cagtgcagtg gtgagaattt ttaatctcatcggcatgatg ctgctcctgt gccactggga 960 tggttgtctt cagttcttag taccactactgcaggacttc ccaccagatt gctgggtgtc 1020 tttaaatgaa atggttaatg attcttggggaaagcagtat tcatacgcac tcttcaaagc 1080 tatgagtcac atgctgtgca ttgggtatggagcccaagcc ccagtcagca tgtctgacct 1140 ctggattacc atgctgagca tgatcgtcggggccacctgc tatgccatgt ttgtcggcca 1200 tgccaccgct ttaatccagt ctctggattcttcgaggcgg cagtatcaag agaagtataa 1260 gcaagtggaa caatacatgt cattccataagttaccagct gatatgcgtc agaagataca 1320 tgattactat gaacacagat accaaggcaaaatctttgat gaggaaaata ttctcaatga 1380 actcaatgat cctctgagag aggagatagtcaacttcaac tgtcggaaac tggtggctac 1440 aatgccttta tttgctaatg cggatcctaattttgtgact gccatgctga gcaagttgag 1500 atttgaggtg tttcaacctg gagattatatcatacgagaa ggagccgtgg gtaaaaaaat 1560 gtatttcatt caacacggtg ttgctggtgtcattacaaaa tccagtaaag aaatgaagct 1620 gacagatggc tcttactttg gagagatttgcctgctgacc aaaggacgtc gtactgccag 1680 tgttcgagct gatacatatt gtcgtctttactcactttcc gtggacaatt tcaacgaggt 1740 cctggaggaa tatccaatga tgaggagagcctttgagaca gttgccattg accgactaga 1800 tcgaatagga aagaaaaatt caattcttctgcaaaagttc cagaaggatc tgaacactgg 1860 tgttttcaac aatcaggaga acgaaatcctcaagcagatt gtgaaacatg acagggagat 1920 ggtgcaggca atcgctccca tcaattatcctcaaatgaca accctgaatt ccacatcgtc 1980 tactacgacc ccgacctccc gcatgaggacacaatctcca ccggtgtaca cagcgaccag 2040 cctgtctcac agcaacctgc actcccccagtcccagcaca cagacccccc agccatcagc 2100 catcctgtca ccctgctcca cgccgaaaaatgaagtgcac aagagcacgc aggcgcttca 2160 caacaccaac ctgacccggg aagtcaggccactctccgcc tcgcagccct cgctgcccca 2220 tgaggtgtcc actctgattt ccagacctcatcccactgtg ggcgagtccc tggcctccat 2280 ccctcaaccc gtgacggcgg tccccggaacgggccttcag gcagggggca ggagcactgt 2340 cccgcagcgc gtcaccctct tccgacagatgtcgtcggga gccatccccc cgaaccgagg 2400 agtccctcca gcaccccctc caccagcagctgctcttcca agagaatctt cctcagtctt 2460 aaacacagac ccagacgcag aaaagccacgatttgcttca aatttatgat ccctgctgat 2520 tgtcaaagca gaaagaaata ctctcataaactgagactat actcagatct tattttattc 2580 tatctcctga tagatccctc tagcctactatgaagagata ttttagacag ctgtggccta 2640 cacgtgaaat gtaaaaatat atatacatatactataaaat atatatctaa attcccaaga 2700 gagggtcaaa agacctgttt agcattcagtgttatatgtc ttcctttctt taaatcatta 2760 aaggatttaa aatgtcaaaa aaaaaaaaaa a2791 2 827 PRT Homo sapiens 2 Met Glu Gly Gly Gly Lys Pro Asn Ser SerSer Asn Ser Arg Asp Asp 1 5 10 15 Gly Asn Ser Val Phe Pro Ala Lys AlaSer Ala Pro Gly Ala Gly Pro 20 25 30 Ala Ala Ala Glu Lys Arg Leu Gly ThrPro Pro Gly Gly Gly Gly Ala 35 40 45 Gly Ala Lys Glu His Gly Asn Ser ValCys Phe Lys Val Asp Gly Gly 50 55 60 Gly Gly Gly Gly Gly Gly Gly Gly GlyGly Glu Glu Pro Ala Gly Gly 65 70 75 80 Phe Glu Asp Ala Glu Gly Pro ArgArg Gln Tyr Gly Phe Met Gln Arg 85 90 95 Gln Phe Thr Ser Met Leu Gln ProGly Val Asn Lys Phe Ser Leu Arg 100 105 110 Met Phe Gly Ser Gln Lys AlaVal Glu Lys Glu Gln Glu Arg Val Lys 115 120 125 Thr Ala Gly Phe Trp IleIle His Pro Tyr Ser Asp Phe Arg Phe Tyr 130 135 140 Trp Asp Leu Ile MetLeu Ile Met Met Val Gly Asn Leu Val Ile Ile 145 150 155 160 Pro Val GlyIle Thr Phe Phe Thr Glu Gln Thr Thr Thr Pro Trp Ile 165 170 175 Ile PheAsn Val Ala Ser Asp Thr Val Phe Leu Leu Asp Leu Ile Met 180 185 190 AsnPhe Arg Thr Gly Thr Val Asn Glu Asp Ser Ser Glu Ile Ile Leu 195 200 205Asp Pro Lys Val Ile Lys Met Asn Tyr Leu Lys Ser Trp Phe Val Val 210 215220 Asp Phe Ile Ser Ser Ile Pro Val Asp Tyr Ile Phe Leu Ile Val Glu 225230 235 240 Lys Gly Met Asp Ser Glu Val Tyr Lys Thr Ala Arg Ala Leu ArgIle 245 250 255 Val Arg Phe Thr Lys Ile Leu Ser Leu Leu Arg Leu Leu ArgLeu Ser 260 265 270 Arg Leu Ile Arg Tyr Ile His Gln Trp Glu Glu Ile PheHis Met Thr 275 280 285 Tyr Asp Leu Ala Ser Ala Val Val Arg Ile Phe AsnLeu Ile Gly Met 290 295 300 Met Leu Leu Leu Cys His Trp Asp Gly Cys LeuGln Phe Leu Val Pro 305 310 315 320 Leu Leu Gln Asp Phe Pro Pro Asp CysTrp Val Ser Leu Asn Glu Met 325 330 335 Val Asn Asp Ser Trp Gly Lys GlnTyr Ser Tyr Ala Leu Phe Lys Ala 340 345 350 Met Ser His Met Leu Cys IleGly Tyr Gly Ala Gln Ala Pro Val Ser 355 360 365 Met Ser Asp Leu Trp IleThr Met Leu Ser Met Ile Val Gly Ala Thr 370 375 380 Cys Tyr Ala Met PheVal Gly His Ala Thr Ala Leu Ile Gln Ser Leu 385 390 395 400 Asp Ser SerArg Arg Gln Tyr Gln Glu Lys Tyr Lys Gln Val Glu Gln 405 410 415 Tyr MetSer Phe His Lys Leu Pro Ala Asp Met Arg Gln Lys Ile His 420 425 430 AspTyr Tyr Glu His Arg Tyr Gln Gly Lys Ile Phe Asp Glu Glu Asn 435 440 445Ile Leu Asn Glu Leu Asn Asp Pro Leu Arg Glu Glu Ile Val Asn Phe 450 455460 Asn Cys Arg Lys Leu Val Ala Thr Met Pro Leu Phe Ala Asn Ala Asp 465470 475 480 Pro Asn Phe Val Thr Ala Met Leu Ser Lys Leu Arg Phe Glu ValPhe 485 490 495 Gln Pro Gly Asp Tyr Ile Ile Arg Glu Gly Ala Val Gly LysLys Met 500 505 510 Tyr Phe Ile Gln His Gly Val Ala Gly Val Ile Thr LysSer Ser Lys 515 520 525 Glu Met Lys Leu Thr Asp Gly Ser Tyr Phe Gly GluIle Cys Leu Leu 530 535 540 Thr Lys Gly Arg Arg Thr Ala Ser Val Arg AlaAsp Thr Tyr Cys Arg 545 550 555 560 Leu Tyr Ser Leu Ser Val Asp Asn PheAsn Glu Val Leu Glu Glu Tyr 565 570 575 Pro Met Met Arg Arg Ala Phe GluThr Val Ala Ile Asp Arg Leu Asp 580 585 590 Arg Ile Gly Lys Lys Asn SerIle Leu Leu Gln Lys Phe Gln Lys Asp 595 600 605 Leu Asn Thr Gly Val PheAsn Asn Gln Glu Asn Glu Ile Leu Lys Gln 610 615 620 Ile Val Lys His AspArg Glu Met Val Gln Ala Ile Ala Pro Ile Asn 625 630 635 640 Tyr Pro GlnMet Thr Thr Leu Asn Ser Thr Ser Ser Thr Thr Thr Pro 645 650 655 Thr SerArg Met Arg Thr Gln Ser Pro Pro Val Tyr Thr Ala Thr Ser 660 665 670 LeuSer His Ser Asn Leu His Ser Pro Ser Pro Ser Thr Gln Thr Pro 675 680 685Gln Pro Ser Ala Ile Leu Ser Pro Cys Ser Thr Pro Lys Asn Glu Val 690 695700 His Lys Ser Thr Gln Ala Leu His Asn Thr Asn Leu Thr Arg Glu Val 705710 715 720 Arg Pro Leu Ser Ala Ser Gln Pro Ser Leu Pro His Glu Val SerThr 725 730 735 Leu Ile Ser Arg Pro His Pro Thr Val Gly Glu Ser Leu AlaSer Ile 740 745 750 Pro Gln Pro Val Thr Ala Val Pro Gly Thr Gly Leu GlnAla Gly Gly 755 760 765 Arg Ser Thr Val Pro Gln Arg Val Thr Leu Phe ArgGln Met Ser Ser 770 775 780 Gly Ala Ile Pro Pro Asn Arg Gly Val Pro ProAla Pro Pro Pro Pro 785 790 795 800 Ala Ala Ala Leu Pro Arg Glu Ser SerSer Val Leu Asn Thr Asp Pro 805 810 815 Asp Ala Glu Lys Pro Arg Phe AlaSer Asn Leu 820 825 3 2980 DNA Homo sapiens 3 ccgtcgccgg ccgcgtcctccgggcatgga aggaggcggc aagcccaact cttcgtctaa 60 cagccgggac gatggcaacagcgtcttccc cgccaaggcg tccgcgccgg gcgcggggcc 120 ggccgcggcc gagaagcgcctgggcacccc gccggggggc ggcggggccg gcgcgaagga 180 gcacggcaac tccgtgtgcttcaaggtgga cggcggtggc ggcggtggcg gcggcggcgg 240 cggcggcgag gagccggcggggggcttcga agacgccgag gggccccggc ggcagtacgg 300 cttcatgcag aggcagttcacctccatgct gcagcccggg gtcaacaaat tctccctccg 360 catgtttggg agccagaaggcggtggaaaa ggagcaggaa agggttaaaa ctgcaggctt 420 ctggattatc cacccttacagtgatttcag gttttactgg gatttaataa tgcttataat 480 gatggttgga aatctagtcatcataccagt tggaatcaca ttctttacag agcaaacaac 540 aacaccatgg attattttcaatgtggcatc agatacagtt ttcctattgg acctgatcat 600 gaattttagg actgggactgtcaatgaaga cagttctgaa atcatcctgg accccaaagt 660 gatcaagatg aattatttaaaaagctggtt tgtggttgac ttcatctcat ccatcccagt 720 ggattatatc tttcttattgtagaaaaagg aatggattct gaagtttaca agacagccag 780 ggcacttcgc attgtgaggtttacaaaaat tctcagtctc ttgcgtttat tacgactttc 840 aaggttaatt agatacatacatcaatggga agagatattc cacatgacat atgatctcgc 900 cagtgcagtg gtgagaatttttaatctcat cggcatgatg ctgctcctgt gccactggga 960 tggttgtctt cagttcttagtaccactact gcaggacttc ccaccagatt gctgggtgtc 1020 tttaaatgaa atggttaatgattcttgggg aaagcagtat tcatacgcac tcttcaaagc 1080 tatgagtcac atgctgtgcattgggtatgg agcccaagcc ccagtcagca tgtctgacct 1140 ctggattacc atgctgagcatgatcgtcgg ggccacctgc tatgccatgt ttgtcggcca 1200 tgccaccgct ttaatccagtctctggattc ttcgaggcgg cagtatcaag agaagtataa 1260 gcaagtggaa caatacatgtcattccataa gttaccagct gatatgcgtc agaagataca 1320 tgattactat gaacacagataccaaggcaa aatctttgat gaggaaaata ttctcaatga 1380 actcaatgat cctctgagagaggagatagt caacttcaac tgtcggaaac tggtggctac 1440 aatgccttta tttgctaatgcggatcctaa ttttgtgact gccatgctga gcaagttgag 1500 atttgaggtg tttcaacctggagattatat catacgagaa ggagccgtgg gtaaaaaaat 1560 gtatttcatt caacacggtgttgctggtgt cattacaaaa tccagtaaag aaatgaagct 1620 gacagatggc tcttactttggagagatttg cctgctgacc aaaggacgtc gtactgccag 1680 tgttcgagct gatacatattgtcgtcttta ctcactttcc gtggacaatt tcaacgaggt 1740 cctggaggaa tatccaatgatgaggagagc ctttgagaca gttgccattg accgactaga 1800 tcgaatagga aagaaaaattcaattcttct gcaaaagttc cagaaggatc tgaacactgg 1860 tgttttcaac aatcaggagaacgaaatcct caagcagatt gtgaaacatg acagggagat 1920 ggtgcaggca atcgctcccatcaattatcc tcaaatgaca accctgaatt ccacatcgtc 1980 tactacgacc ccgacctcccgcatgaggac acaatctcca ccggtgtaca cagcgaccag 2040 cctgtctcac agcaacctgcactcccccag tcccagcaca cagacccccc agccatcagc 2100 catcctgtca ccctgctcctacaccaccgc ggtctgcagc cctcctgtac agagccctct 2160 ggccgctcga actttccactatgcctcccc caccgcctcc cagctgtcac tcatgcaaca 2220 gcagccgcag cagcaggtacagcagtccca gccgccgcag actcagccac agcagccgtc 2280 cccgcagcca cagacacctggcagctccac gccgaaaaat gaagtgcaca agagcacgca 2340 ggcgcttcac aacaccaacctgacccggga agtcaggcca ctctccgcct cgcagccctc 2400 gctgccccat gaggtgtccactctgatttc cagacctcat cccactgtgg gcgagtccct 2460 ggcctccatc cctcaacccgtgacggcggt ccccggaacg ggccttcagg cagggggcag 2520 gagcactgtc ccgcagcgcgtcaccctctt ccgacagatg tcgtcgggag ccatcccccc 2580 gaaccgagga gtccctccagcaccccctcc accagcagct gctcttccaa gagaatcttc 2640 ctcagtctta aacacagacccagacgcaga aaagccacga tttgcttcaa atttatgatc 2700 cctgctgatt gtcaaagcagaaagaaatac tctcataaac tgagactata ctcagatctt 2760 attttattct atctcctgatagatccctct agcctactat gaagagatat tttagacagc 2820 tgtggcctac acgtgaaatgtaaaaatata tatacatata ctataaaata tatatctaaa 2880 ttcccaagag agggtcaaaagacctgttta gcattcagtg ttatatgtct tcctttcttt 2940 aaatcattaa aggatttaaaatgtcaaaaa aaaaaaaaaa 2980 4 890 PRT Homo sapiens 4 Met Glu Gly Gly GlyLys Pro Asn Ser Ser Ser Asn Ser Arg Asp Asp 1 5 10 15 Gly Asn Ser ValPhe Pro Ala Lys Ala Ser Ala Pro Gly Ala Gly Pro 20 25 30 Ala Ala Ala GluLys Arg Leu Gly Thr Pro Pro Gly Gly Gly Gly Ala 35 40 45 Gly Ala Lys GluHis Gly Asn Ser Val Cys Phe Lys Val Asp Gly Gly 50 55 60 Gly Gly Gly GlyGly Gly Gly Gly Gly Gly Glu Glu Pro Ala Gly Gly 65 70 75 80 Phe Glu AspAla Glu Gly Pro Arg Arg Gln Tyr Gly Phe Met Gln Arg 85 90 95 Gln Phe ThrSer Met Leu Gln Pro Gly Val Asn Lys Phe Ser Leu Arg 100 105 110 Met PheGly Ser Gln Lys Ala Val Glu Lys Glu Gln Glu Arg Val Lys 115 120 125 ThrAla Gly Phe Trp Ile Ile His Pro Tyr Ser Asp Phe Arg Phe Tyr 130 135 140Trp Asp Leu Ile Met Leu Ile Met Met Val Gly Asn Leu Val Ile Ile 145 150155 160 Pro Val Gly Ile Thr Phe Phe Thr Glu Gln Thr Thr Thr Pro Trp Ile165 170 175 Ile Phe Asn Val Ala Ser Asp Thr Val Phe Leu Leu Asp Leu IleMet 180 185 190 Asn Phe Arg Thr Gly Thr Val Asn Glu Asp Ser Ser Glu IleIle Leu 195 200 205 Asp Pro Lys Val Ile Lys Met Asn Tyr Leu Lys Ser TrpPhe Val Val 210 215 220 Asp Phe Ile Ser Ser Ile Pro Val Asp Tyr Ile PheLeu Ile Val Glu 225 230 235 240 Lys Gly Met Asp Ser Glu Val Tyr Lys ThrAla Arg Ala Leu Arg Ile 245 250 255 Val Arg Phe Thr Lys Ile Leu Ser LeuLeu Arg Leu Leu Arg Leu Ser 260 265 270 Arg Leu Ile Arg Tyr Ile His GlnTrp Glu Glu Ile Phe His Met Thr 275 280 285 Tyr Asp Leu Ala Ser Ala ValVal Arg Ile Phe Asn Leu Ile Gly Met 290 295 300 Met Leu Leu Leu Cys HisTrp Asp Gly Cys Leu Gln Phe Leu Val Pro 305 310 315 320 Leu Leu Gln AspPhe Pro Pro Asp Cys Trp Val Ser Leu Asn Glu Met 325 330 335 Val Asn AspSer Trp Gly Lys Gln Tyr Ser Tyr Ala Leu Phe Lys Ala 340 345 350 Met SerHis Met Leu Cys Ile Gly Tyr Gly Ala Gln Ala Pro Val Ser 355 360 365 MetSer Asp Leu Trp Ile Thr Met Leu Ser Met Ile Val Gly Ala Thr 370 375 380Cys Tyr Ala Met Phe Val Gly His Ala Thr Ala Leu Ile Gln Ser Leu 385 390395 400 Asp Ser Ser Arg Arg Gln Tyr Gln Glu Lys Tyr Lys Gln Val Glu Gln405 410 415 Tyr Met Ser Phe His Lys Leu Pro Ala Asp Met Arg Gln Lys IleHis 420 425 430 Asp Tyr Tyr Glu His Arg Tyr Gln Gly Lys Ile Phe Asp GluGlu Asn 435 440 445 Ile Leu Asn Glu Leu Asn Asp Pro Leu Arg Glu Glu IleVal Asn Phe 450 455 460 Asn Cys Arg Lys Leu Val Ala Thr Met Pro Leu PheAla Asn Ala Asp 465 470 475 480 Pro Asn Phe Val Thr Ala Met Leu Ser LysLeu Arg Phe Glu Val Phe 485 490 495 Gln Pro Gly Asp Tyr Ile Ile Arg GluGly Ala Val Gly Lys Lys Met 500 505 510 Tyr Phe Ile Gln His Gly Val AlaGly Val Ile Thr Lys Ser Ser Lys 515 520 525 Glu Met Lys Leu Thr Asp GlySer Tyr Phe Gly Glu Ile Cys Leu Leu 530 535 540 Thr Lys Gly Arg Arg ThrAla Ser Val Arg Ala Asp Thr Tyr Cys Arg 545 550 555 560 Leu Tyr Ser LeuSer Val Asp Asn Phe Asn Glu Val Leu Glu Glu Tyr 565 570 575 Pro Met MetArg Arg Ala Phe Glu Thr Val Ala Ile Asp Arg Leu Asp 580 585 590 Arg IleGly Lys Lys Asn Ser Ile Leu Leu Gln Lys Phe Gln Lys Asp 595 600 605 LeuAsn Thr Gly Val Phe Asn Asn Gln Glu Asn Glu Ile Leu Lys Gln 610 615 620Ile Val Lys His Asp Arg Glu Met Val Gln Ala Ile Ala Pro Ile Asn 625 630635 640 Tyr Pro Gln Met Thr Thr Leu Asn Ser Thr Ser Ser Thr Thr Thr Pro645 650 655 Thr Ser Arg Met Arg Thr Gln Ser Pro Pro Val Tyr Thr Ala ThrSer 660 665 670 Leu Ser His Ser Asn Leu His Ser Pro Ser Pro Ser Thr GlnThr Pro 675 680 685 Gln Pro Ser Ala Ile Leu Ser Pro Cys Ser Tyr Thr ThrAla Val Cys 690 695 700 Ser Pro Pro Val Gln Ser Pro Leu Ala Ala Arg ThrPhe His Tyr Ala 705 710 715 720 Ser Pro Thr Ala Ser Gln Leu Ser Leu MetGln Gln Gln Pro Gln Gln 725 730 735 Gln Val Gln Gln Ser Gln Pro Pro GlnThr Gln Pro Gln Gln Pro Ser 740 745 750 Pro Gln Pro Gln Thr Pro Gly SerSer Thr Pro Lys Asn Glu Val His 755 760 765 Lys Ser Thr Gln Ala Leu HisAsn Thr Asn Leu Thr Arg Glu Val Arg 770 775 780 Pro Leu Ser Ala Ser GlnPro Ser Leu Pro His Glu Val Ser Thr Leu 785 790 795 800 Ile Ser Arg ProHis Pro Thr Val Gly Glu Ser Leu Ala Ser Ile Pro 805 810 815 Gln Pro ValThr Ala Val Pro Gly Thr Gly Leu Gln Ala Gly Gly Arg 820 825 830 Ser ThrVal Pro Gln Arg Val Thr Leu Phe Arg Gln Met Ser Ser Gly 835 840 845 AlaIle Pro Pro Asn Arg Gly Val Pro Pro Ala Pro Pro Pro Pro Ala 850 855 860Ala Ala Leu Pro Arg Glu Ser Ser Ser Val Leu Asn Thr Asp Pro Asp 865 870875 880 Ala Glu Lys Pro Arg Phe Ala Ser Asn Leu 885 890 5 2791 DNA Homosapiens 5 ccgtcgccgg ccgcgtcctc cgggcatgga aggaggcggc aagcccaactcttcgtctaa 60 cagccgggac gatggcaaca gcgtcttccc cgccaaggcg tccgcgacgggcgcggggcc 120 ggccgcggcc gagaagcgcc tgggcacccc gccggggggc ggcggggccggcgcgaagga 180 gcacggcaac tccgtgtgct tcaaggtgga cggcggtggc ggcggtggcggcggcggcgg 240 cggcggcgag gagccggcgg ggggcttcga agacgccgag gggccccggcggcagtacgg 300 cttcatgcag aggcagttca cctccatgct gcagcccggg gtcaacaaattctccctccg 360 catgtttggg agccagaagg cggtggaaaa ggagcaggaa agggttaaaactgcaggctt 420 ctggattatc cacccttaca gtgatttcag gttttactgg gatttaataatgcttataat 480 gatggttgga aatctagtca tcataccagt tggaatcaca ttctttacagagcaaacaac 540 aacaccatgg attattttca atgtggcatc agatacagtt ttcctattggacctgatcat 600 gaattttagg actgggactg tcaatgaaga cagttctgaa atcatcctggaccccaaagt 660 gatcaagatg aattatttaa aaagctggtt tgtggttgac ttcatctcatccatcccagt 720 ggattatatc tttcttattg tagaaaaagg aatggattct gaagtttacaagacagccag 780 ggcacttcgc attgtgaggt ttacaaaaat tctcagtctc ttgcgtttattacgactttc 840 aaggttaatt agatacatac atcaatggga agagatattc cacatgacatatgatctcgc 900 cagtgcagtg gtgagaattt ttaatctcat cggcatgatg ctgctcctgtgccactggga 960 tggttgtctt cagttcttag taccactact gcaggacttc ccaccagattgctgggtgtc 1020 tttaaatgaa atggttaatg attcttgggg aaagcagtat tcatacgcactcttcaaagc 1080 tatgagtcac atgctgtgca ttgggtatgg agcccaagcc ccagtcagcatgtctgacct 1140 ctggattacc atgctgagca tgatcgtcgg ggccacctgc tatgccatgtttgtcggcca 1200 tgccaccgct ttaatccagt ctctggattc ttcgaggcgg cagtatcaagagaagtataa 1260 gcaagtggaa caatacatgt cattccataa gttaccagct gatatgcgtcagaagataca 1320 tgattactat gaacacagat accaaggcaa aatctttgat gaggaaaatattctcaatga 1380 actcaatgat cctctgagag aggagatagt caacttcaac tgtcggaaactggtggctac 1440 aatgccttta tttgctaatg cggatcctaa ttttgtgact gccatgctgagcaagttgag 1500 atttgaggtg tttcaacctg gagattatat catacgagaa ggagccgtgggtaaaaaaat 1560 gtatttcatt caacacggtg ttgctggtgt cattacaaaa tccagtaaagaaatgaagct 1620 gacagatggc tcttactttg gagagatttg cctgctgacc aaaggacgtcgtactgccag 1680 tgttcgagct gatacatatt gtcgtcttta ctcactttcc gtggacaatttcaacgaggt 1740 cctggaggaa tatccaatga tgaggagagc ctttgagaca gttgccattgaccgactaga 1800 tcgaatagga aagaaaaatt caattcttct gcaaaagttc cagaaggatctgaacactgg 1860 tgttttcaac aatcaggaga acgaaatcct caagcagatt gtgaaacatgacagggagat 1920 ggtgcaggca atcgctccca tcaattatcc tcaaatgaca accctgaattccacatcgtc 1980 tactacgacc ccgacctccc gcatgaggac acaatctcca ccggtgtacacagcgaccag 2040 cctgtctcac agcaacctgc acttccccag tcccagcaca cagaccccccagccatcagc 2100 catcctgtca ccctgctcca cgccgaaaaa tgaagtgcac aagagcacgcaggcgcttca 2160 caacaccaac ctgacccggg aagtcaggcc actctccgcc tcgcagccctcgctgcccca 2220 tgaggtgtcc actctgattt ccagacctca tcccactgtg ggcgagtccctggcctccat 2280 ccctcaaccc gtgacggcgg tccccggaac gggccttcag gcagggggcaggagcactgt 2340 cccgcagcgc gtcaccctct tccgacagat gtcgtcggga gccatccccccgaaccgagg 2400 agtccctcca gcaccccctc caccagcagc tgctcttcca agagaatcttcctcagtctt 2460 aaacacagac ccagacgcag aaaagccacg atttgcttca aatttatgatccctgctgat 2520 tgtcaaagca gaaagaaata ctctcataaa ctgagactat actcagatcttattttattc 2580 tatctcctga tagatccctc tagcctacta tgaagagata ttttagacagctgtggccta 2640 cacgtgaaat gtaaaaatat atatacatat actataaaat atatatctaaattcccaaga 2700 gagggtcaaa agacctgttt agcattcagt gttatatgtc ttcctttctttaaatcatta 2760 aaggatttaa aatgtcaaaa aaaaaaaaaa a 2791 6 827 PRT Homosapiens 6 Met Glu Gly Gly Gly Lys Pro Asn Ser Ser Ser Asn Ser Arg AspAsp 1 5 10 15 Gly Asn Ser Val Phe Pro Ala Lys Ala Ser Ala Thr Gly AlaGly Pro 20 25 30 Ala Ala Ala Glu Lys Arg Leu Gly Thr Pro Pro Gly Gly GlyGly Ala 35 40 45 Gly Ala Lys Glu His Gly Asn Ser Val Cys Phe Lys Val AspGly Gly 50 55 60 Gly Gly Gly Gly Gly Gly Gly Gly Gly Gly Glu Glu Pro AlaGly Gly 65 70 75 80 Phe Glu Asp Ala Glu Gly Pro Arg Arg Gln Tyr Gly PheMet Gln Arg 85 90 95 Gln Phe Thr Ser Met Leu Gln Pro Gly Val Asn Lys PheSer Leu Arg 100 105 110 Met Phe Gly Ser Gln Lys Ala Val Glu Lys Glu GlnGlu Arg Val Lys 115 120 125 Thr Ala Gly Phe Trp Ile Ile His Pro Tyr SerAsp Phe Arg Phe Tyr 130 135 140 Trp Asp Leu Ile Met Leu Ile Met Met ValGly Asn Leu Val Ile Ile 145 150 155 160 Pro Val Gly Ile Thr Phe Phe ThrGlu Gln Thr Thr Thr Pro Trp Ile 165 170 175 Ile Phe Asn Val Ala Ser AspThr Val Phe Leu Leu Asp Leu Ile Met 180 185 190 Asn Phe Arg Thr Gly ThrVal Asn Glu Asp Ser Ser Glu Ile Ile Leu 195 200 205 Asp Pro Lys Val IleLys Met Asn Tyr Leu Lys Ser Trp Phe Val Val 210 215 220 Asp Phe Ile SerSer Ile Pro Val Asp Tyr Ile Phe Leu Ile Val Glu 225 230 235 240 Lys GlyMet Asp Ser Glu Val Tyr Lys Thr Ala Arg Ala Leu Arg Ile 245 250 255 ValArg Phe Thr Lys Ile Leu Ser Leu Leu Arg Leu Leu Arg Leu Ser 260 265 270Arg Leu Ile Arg Tyr Ile His Gln Trp Glu Glu Ile Phe His Met Thr 275 280285 Tyr Asp Leu Ala Ser Ala Val Val Arg Ile Phe Asn Leu Ile Gly Met 290295 300 Met Leu Leu Leu Cys His Trp Asp Gly Cys Leu Gln Phe Leu Val Pro305 310 315 320 Leu Leu Gln Asp Phe Pro Pro Asp Cys Trp Val Ser Leu AsnGlu Met 325 330 335 Val Asn Asp Ser Trp Gly Lys Gln Tyr Ser Tyr Ala LeuPhe Lys Ala 340 345 350 Met Ser His Met Leu Cys Ile Gly Tyr Gly Ala GlnAla Pro Val Ser 355 360 365 Met Ser Asp Leu Trp Ile Thr Met Leu Ser MetIle Val Gly Ala Thr 370 375 380 Cys Tyr Ala Met Phe Val Gly His Ala ThrAla Leu Ile Gln Ser Leu 385 390 395 400 Asp Ser Ser Arg Arg Gln Tyr GlnGlu Lys Tyr Lys Gln Val Glu Gln 405 410 415 Tyr Met Ser Phe His Lys LeuPro Ala Asp Met Arg Gln Lys Ile His 420 425 430 Asp Tyr Tyr Glu His ArgTyr Gln Gly Lys Ile Phe Asp Glu Glu Asn 435 440 445 Ile Leu Asn Glu LeuAsn Asp Pro Leu Arg Glu Glu Ile Val Asn Phe 450 455 460 Asn Cys Arg LysLeu Val Ala Thr Met Pro Leu Phe Ala Asn Ala Asp 465 470 475 480 Pro AsnPhe Val Thr Ala Met Leu Ser Lys Leu Arg Phe Glu Val Phe 485 490 495 GlnPro Gly Asp Tyr Ile Ile Arg Glu Gly Ala Val Gly Lys Lys Met 500 505 510Tyr Phe Ile Gln His Gly Val Ala Gly Val Ile Thr Lys Ser Ser Lys 515 520525 Glu Met Lys Leu Thr Asp Gly Ser Tyr Phe Gly Glu Ile Cys Leu Leu 530535 540 Thr Lys Gly Arg Arg Thr Ala Ser Val Arg Ala Asp Thr Tyr Cys Arg545 550 555 560 Leu Tyr Ser Leu Ser Val Asp Asn Phe Asn Glu Val Leu GluGlu Tyr 565 570 575 Pro Met Met Arg Arg Ala Phe Glu Thr Val Ala Ile AspArg Leu Asp 580 585 590 Arg Ile Gly Lys Lys Asn Ser Ile Leu Leu Gln LysPhe Gln Lys Asp 595 600 605 Leu Asn Thr Gly Val Phe Asn Asn Gln Glu AsnGlu Ile Leu Lys Gln 610 615 620 Ile Val Lys His Asp Arg Glu Met Val GlnAla Ile Ala Pro Ile Asn 625 630 635 640 Tyr Pro Gln Met Thr Thr Leu AsnSer Thr Ser Ser Thr Thr Thr Pro 645 650 655 Thr Ser Arg Met Arg Thr GlnSer Pro Pro Val Tyr Thr Ala Thr Ser 660 665 670 Leu Ser His Ser Asn LeuHis Phe Pro Ser Pro Ser Thr Gln Thr Pro 675 680 685 Gln Pro Ser Ala IleLeu Ser Pro Cys Ser Thr Pro Lys Asn Glu Val 690 695 700 His Lys Ser ThrGln Ala Leu His Asn Thr Asn Leu Thr Arg Glu Val 705 710 715 720 Arg ProLeu Ser Ala Ser Gln Pro Ser Leu Pro His Glu Val Ser Thr 725 730 735 LeuIle Ser Arg Pro His Pro Thr Val Gly Glu Ser Leu Ala Ser Ile 740 745 750Pro Gln Pro Val Thr Ala Val Pro Gly Thr Gly Leu Gln Ala Gly Gly 755 760765 Arg Ser Thr Val Pro Gln Arg Val Thr Leu Phe Arg Gln Met Ser Ser 770775 780 Gly Ala Ile Pro Pro Asn Arg Gly Val Pro Pro Ala Pro Pro Pro Pro785 790 795 800 Ala Ala Ala Leu Pro Arg Glu Ser Ser Ser Val Leu Asn ThrAsp Pro 805 810 815 Asp Ala Glu Lys Pro Arg Phe Ala Ser Asn Leu 820 8257 2980 DNA Homo sapiens 7 ccgtcgccgg ccgcgtcctc cgggcatgga aggaggcggcaagcccaact cttcgtctaa 60 cagccgggac gatggcaaca gcgtcttccc cgccaaggcgtccgcgacgg gcgcggggcc 120 ggccgcggcc gagaagcgcc tgggcacccc gccggggggcggcggggccg gcgcgaagga 180 gcacggcaac tccgtgtgct tcaaggtgga cggcggtggcggcggtggcg gcggcggcgg 240 cggcggcgag gagccggcgg ggggcttcga agacgccgaggggccccggc ggcagtacgg 300 cttcatgcag aggcagttca cctccatgct gcagcccggggtcaacaaat tctccctccg 360 catgtttggg agccagaagg cggtggaaaa ggagcaggaaagggttaaaa ctgcaggctt 420 ctggattatc cacccttaca gtgatttcag gttttactgggatttaataa tgcttataat 480 gatggttgga aatctagtca tcataccagt tggaatcacattctttacag agcaaacaac 540 aacaccatgg attattttca atgtggcatc agatacagttttcctattgg acctgatcat 600 gaattttagg actgggactg tcaatgaaga cagttctgaaatcatcctgg accccaaagt 660 gatcaagatg aattatttaa aaagctggtt tgtggttgacttcatctcat ccatcccagt 720 ggattatatc tttcttattg tagaaaaagg aatggattctgaagtttaca agacagccag 780 ggcacttcgc attgtgaggt ttacaaaaat tctcagtctcttgcgtttat tacgactttc 840 aaggttaatt agatacatac atcaatggga agagatattccacatgacat atgatctcgc 900 cagtgcagtg gtgagaattt ttaatctcat cggcatgatgctgctcctgt gccactggga 960 tggttgtctt cagttcttag taccactact gcaggacttcccaccagatt gctgggtgtc 1020 tttaaatgaa atggttaatg attcttgggg aaagcagtattcatacgcac tcttcaaagc 1080 tatgagtcac atgctgtgca ttgggtatgg agcccaagccccagtcagca tgtctgacct 1140 ctggattacc atgctgagca tgatcgtcgg ggccacctgctatgccatgt ttgtcggcca 1200 tgccaccgct ttaatccagt ctctggattc ttcgaggcggcagtatcaag agaagtataa 1260 gcaagtggaa caatacatgt cattccataa gttaccagctgatatgcgtc agaagataca 1320 tgattactat gaacacagat accaaggcaa aatctttgatgaggaaaata ttctcaatga 1380 actcaatgat cctctgagag aggagatagt caacttcaactgtcggaaac tggtggctac 1440 aatgccttta tttgctaatg cggatcctaa ttttgtgactgccatgctga gcaagttgag 1500 atttgaggtg tttcaacctg gagattatat catacgagaaggagccgtgg gtaaaaaaat 1560 gtatttcatt caacacggtg ttgctggtgt cattacaaaatccagtaaag aaatgaagct 1620 gacagatggc tcttactttg gagagatttg cctgctgaccaaaggacgtc gtactgccag 1680 tgttcgagct gatacatatt gtcgtcttta ctcactttccgtggacaatt tcaacgaggt 1740 cctggaggaa tatccaatga tgaggagagc ctttgagacagttgccattg accgactaga 1800 tcgaatagga aagaaaaatt caattcttct gcaaaagttccagaaggatc tgaacactgg 1860 tgttttcaac aatcaggaga acgaaatcct caagcagattgtgaaacatg acagggagat 1920 ggtgcaggca atcgctccca tcaattatcc tcaaatgacaaccctgaatt ccacatcgtc 1980 tactacgacc ccgacctccc gcatgaggac acaatctccaccggtgtaca cagcgaccag 2040 cctgtctcac agcaacctgc acttccccag tcccagcacacagacccccc agccatcagc 2100 catcctgtca ccctgctcct acaccaccgc ggtctgcagccctcctgtac agagccctct 2160 ggccgctcga actttccact atgcctcccc caccgcctcccagctgtcac tcatgcaaca 2220 gcagccgcag cagcaggtac agcagtccca gccgccgcagactcagccac agcagccgtc 2280 cccgcagcca cagacacctg gcagctccac gccgaaaaatgaagtgcaca agagcacgca 2340 ggcgcttcac aacaccaacc tgacccggga agtcaggccactctccgcct cgcagccctc 2400 gctgccccat gaggtgtcca ctctgatttc cagacctcatcccactgtgg gcgagtccct 2460 ggcctccatc cctcaacccg tgacggcggt ccccggaacgggccttcagg cagggggcag 2520 gagcactgtc ccgcagcgcg tcaccctctt ccgacagatgtcgtcgggag ccatcccccc 2580 gaaccgagga gtccctccag caccccctcc accagcagctgctcttccaa gagaatcttc 2640 ctcagtctta aacacagacc cagacgcaga aaagccacgatttgcttcaa atttatgatc 2700 cctgctgatt gtcaaagcag aaagaaatac tctcataaactgagactata ctcagatctt 2760 attttattct atctcctgat agatccctct agcctactatgaagagatat tttagacagc 2820 tgtggcctac acgtgaaatg taaaaatata tatacatatactataaaata tatatctaaa 2880 ttcccaagag agggtcaaaa gacctgttta gcattcagtgttatatgtct tcctttcttt 2940 aaatcattaa aggatttaaa atgtcaaaaa aaaaaaaaaa2980 8 890 PRT Homo sapiens 8 Met Glu Gly Gly Gly Lys Pro Asn Ser SerSer Asn Ser Arg Asp Asp 1 5 10 15 Gly Asn Ser Val Phe Pro Ala Lys AlaSer Ala Thr Gly Ala Gly Pro 20 25 30 Ala Ala Ala Glu Lys Arg Leu Gly ThrPro Pro Gly Gly Gly Gly Ala 35 40 45 Gly Ala Lys Glu His Gly Asn Ser ValCys Phe Lys Val Asp Gly Gly 50 55 60 Gly Gly Gly Gly Gly Gly Gly Gly GlyGly Glu Glu Pro Ala Gly Gly 65 70 75 80 Phe Glu Asp Ala Glu Gly Pro ArgArg Gln Tyr Gly Phe Met Gln Arg 85 90 95 Gln Phe Thr Ser Met Leu Gln ProGly Val Asn Lys Phe Ser Leu Arg 100 105 110 Met Phe Gly Ser Gln Lys AlaVal Glu Lys Glu Gln Glu Arg Val Lys 115 120 125 Thr Ala Gly Phe Trp IleIle His Pro Tyr Ser Asp Phe Arg Phe Tyr 130 135 140 Trp Asp Leu Ile MetLeu Ile Met Met Val Gly Asn Leu Val Ile Ile 145 150 155 160 Pro Val GlyIle Thr Phe Phe Thr Glu Gln Thr Thr Thr Pro Trp Ile 165 170 175 Ile PheAsn Val Ala Ser Asp Thr Val Phe Leu Leu Asp Leu Ile Met 180 185 190 AsnPhe Arg Thr Gly Thr Val Asn Glu Asp Ser Ser Glu Ile Ile Leu 195 200 205Asp Pro Lys Val Ile Lys Met Asn Tyr Leu Lys Ser Trp Phe Val Val 210 215220 Asp Phe Ile Ser Ser Ile Pro Val Asp Tyr Ile Phe Leu Ile Val Glu 225230 235 240 Lys Gly Met Asp Ser Glu Val Tyr Lys Thr Ala Arg Ala Leu ArgIle 245 250 255 Val Arg Phe Thr Lys Ile Leu Ser Leu Leu Arg Leu Leu ArgLeu Ser 260 265 270 Arg Leu Ile Arg Tyr Ile His Gln Trp Glu Glu Ile PheHis Met Thr 275 280 285 Tyr Asp Leu Ala Ser Ala Val Val Arg Ile Phe AsnLeu Ile Gly Met 290 295 300 Met Leu Leu Leu Cys His Trp Asp Gly Cys LeuGln Phe Leu Val Pro 305 310 315 320 Leu Leu Gln Asp Phe Pro Pro Asp CysTrp Val Ser Leu Asn Glu Met 325 330 335 Val Asn Asp Ser Trp Gly Lys GlnTyr Ser Tyr Ala Leu Phe Lys Ala 340 345 350 Met Ser His Met Leu Cys IleGly Tyr Gly Ala Gln Ala Pro Val Ser 355 360 365 Met Ser Asp Leu Trp IleThr Met Leu Ser Met Ile Val Gly Ala Thr 370 375 380 Cys Tyr Ala Met PheVal Gly His Ala Thr Ala Leu Ile Gln Ser Leu 385 390 395 400 Asp Ser SerArg Arg Gln Tyr Gln Glu Lys Tyr Lys Gln Val Glu Gln 405 410 415 Tyr MetSer Phe His Lys Leu Pro Ala Asp Met Arg Gln Lys Ile His 420 425 430 AspTyr Tyr Glu His Arg Tyr Gln Gly Lys Ile Phe Asp Glu Glu Asn 435 440 445Ile Leu Asn Glu Leu Asn Asp Pro Leu Arg Glu Glu Ile Val Asn Phe 450 455460 Asn Cys Arg Lys Leu Val Ala Thr Met Pro Leu Phe Ala Asn Ala Asp 465470 475 480 Pro Asn Phe Val Thr Ala Met Leu Ser Lys Leu Arg Phe Glu ValPhe 485 490 495 Gln Pro Gly Asp Tyr Ile Ile Arg Glu Gly Ala Val Gly LysLys Met 500 505 510 Tyr Phe Ile Gln His Gly Val Ala Gly Val Ile Thr LysSer Ser Lys 515 520 525 Glu Met Lys Leu Thr Asp Gly Ser Tyr Phe Gly GluIle Cys Leu Leu 530 535 540 Thr Lys Gly Arg Arg Thr Ala Ser Val Arg AlaAsp Thr Tyr Cys Arg 545 550 555 560 Leu Tyr Ser Leu Ser Val Asp Asn PheAsn Glu Val Leu Glu Glu Tyr 565 570 575 Pro Met Met Arg Arg Ala Phe GluThr Val Ala Ile Asp Arg Leu Asp 580 585 590 Arg Ile Gly Lys Lys Asn SerIle Leu Leu Gln Lys Phe Gln Lys Asp 595 600 605 Leu Asn Thr Gly Val PheAsn Asn Gln Glu Asn Glu Ile Leu Lys Gln 610 615 620 Ile Val Lys His AspArg Glu Met Val Gln Ala Ile Ala Pro Ile Asn 625 630 635 640 Tyr Pro GlnMet Thr Thr Leu Asn Ser Thr Ser Ser Thr Thr Thr Pro 645 650 655 Thr SerArg Met Arg Thr Gln Ser Pro Pro Val Tyr Thr Ala Thr Ser 660 665 670 LeuSer His Ser Asn Leu His Phe Pro Ser Pro Ser Thr Gln Thr Pro 675 680 685Gln Pro Ser Ala Ile Leu Ser Pro Cys Ser Tyr Thr Thr Ala Val Cys 690 695700 Ser Pro Pro Val Gln Ser Pro Leu Ala Ala Arg Thr Phe His Tyr Ala 705710 715 720 Ser Pro Thr Ala Ser Gln Leu Ser Leu Met Gln Gln Gln Pro GlnGln 725 730 735 Gln Val Gln Gln Ser Gln Pro Pro Gln Thr Gln Pro Gln GlnPro Ser 740 745 750 Pro Gln Pro Gln Thr Pro Gly Ser Ser Thr Pro Lys AsnGlu Val His 755 760 765 Lys Ser Thr Gln Ala Leu His Asn Thr Asn Leu ThrArg Glu Val Arg 770 775 780 Pro Leu Ser Ala Ser Gln Pro Ser Leu Pro HisGlu Val Ser Thr Leu 785 790 795 800 Ile Ser Arg Pro His Pro Thr Val GlyGlu Ser Leu Ala Ser Ile Pro 805 810 815 Gln Pro Val Thr Ala Val Pro GlyThr Gly Leu Gln Ala Gly Gly Arg 820 825 830 Ser Thr Val Pro Gln Arg ValThr Leu Phe Arg Gln Met Ser Ser Gly 835 840 845 Ala Ile Pro Pro Asn ArgGly Val Pro Pro Ala Pro Pro Pro Pro Ala 850 855 860 Ala Ala Leu Pro ArgGlu Ser Ser Ser Val Leu Asn Thr Asp Pro Asp 865 870 875 880 Ala Glu LysPro Arg Phe Ala Ser Asn Leu 885 890 9 1046 DNA Homo sapiens 9 cgagtaattttgtgactgcc atgctgagca agttgagatt tgaggtgttt caacctggag 60 attatatcatacgagaagga gccgtgggta aaaaaatgta tttcattcaa cacggtgttg 120 ctggtgtcattacaaaatcc agtaaagaaa tgaagctgac agatggctct tactttggag 180 agatttgcctgctgaccaaa ggacgtcgta ctgccagtgt tcgagctgat acatattgtc 240 gtctttactcactttccgtg gacaatttca acgaggtcct ggaggaatat ccaatgatga 300 ggagagcctttgagacagtt gccattgacc gactagatcg aataggaaag aaaaattcaa 360 ttcttctgcaaaagttccag aaggatctga acactggtgt tttcaacaat caggagaacg 420 aaatcctcaagcagattgtg aaacatgaca gggagatggt gcaggcaatc gctcccatca 480 attatcctcaaatgacaacc ctgaattcca catcgtctac tacgaccccg acctcccgca 540 tgaggacacaatctccaccg gtgtacacag cgaccagcct gtctcacagc aacctgcact 600 cccccagtcccagcacacag accccccagc catcagccat cctgtcaccc tgctccacgc 660 cgaaaaatgaagtgcacaag agcacgcagg cgcttcacaa caccaacctg acccgggaag 720 tcaggccactctccgcctcg cagccctcgc tgccccatga ggtgtccact ctgatttcca 780 gacctcatcccactgtgggc gagtccctgg cctccatccc tcaacccgtg acggcggtcc 840 ccggaacgggccttcaggca gggggcagga gcactgtccc gcagcgcgtc accctcttcc 900 gacagatgtcgtcgggagcc atccccccga accgaggagt ccctccagca ccccctccac 960 cagcagctgctcttccaaga gaatcttcct cagtcttaaa cacagaccca gacgcagaaa 1020 agccacgatttgcttcaaat ttatga 1046 10 346 PRT Homo sapiens 10 Asn Phe Val Thr AlaMet Leu Ser Lys Leu Arg Phe Glu Val Phe Gln 1 5 10 15 Pro Gly Asp TyrIle Ile Arg Glu Gly Ala Val Gly Lys Lys Met Tyr 20 25 30 Phe Ile Gln HisGly Val Ala Gly Val Ile Thr Lys Ser Ser Lys Glu 35 40 45 Met Lys Leu ThrAsp Gly Ser Tyr Phe Gly Glu Ile Cys Leu Leu Thr 50 55 60 Lys Gly Arg ArgThr Ala Ser Val Arg Ala Asp Thr Tyr Cys Arg Leu 65 70 75 80 Tyr Ser LeuSer Val Asp Asn Phe Asn Glu Val Leu Glu Glu Tyr Pro 85 90 95 Met Met ArgArg Ala Phe Glu Thr Val Ala Ile Asp Arg Leu Asp Arg 100 105 110 Ile GlyLys Lys Asn Ser Ile Leu Leu Gln Lys Phe Gln Lys Asp Leu 115 120 125 AsnThr Gly Val Phe Asn Asn Gln Glu Asn Glu Ile Leu Lys Gln Ile 130 135 140Val Lys His Asp Arg Glu Met Val Gln Ala Ile Ala Pro Ile Asn Tyr 145 150155 160 Pro Gln Met Thr Thr Leu Asn Ser Thr Ser Ser Thr Thr Thr Pro Thr165 170 175 Ser Arg Met Arg Thr Gln Ser Pro Pro Val Tyr Thr Ala Thr SerLeu 180 185 190 Ser His Ser Asn Leu His Ser Pro Ser Pro Ser Thr Gln ThrPro Gln 195 200 205 Pro Ser Ala Ile Leu Ser Pro Cys Ser Thr Pro Lys AsnGlu Val His 210 215 220 Lys Ser Thr Gln Ala Leu His Asn Thr Asn Leu ThrArg Glu Val Arg 225 230 235 240 Pro Leu Ser Ala Ser Gln Pro Ser Leu ProHis Glu Val Ser Thr Leu 245 250 255 Ile Ser Arg Pro His Pro Thr Val GlyGlu Ser Leu Ala Ser Ile Pro 260 265 270 Gln Pro Val Thr Ala Val Pro GlyThr Gly Leu Gln Ala Gly Gly Arg 275 280 285 Ser Thr Val Pro Gln Arg ValThr Leu Phe Arg Gln Met Ser Ser Gly 290 295 300 Ala Ile Pro Pro Asn ArgGly Val Pro Pro Ala Pro Pro Pro Pro Ala 305 310 315 320 Ala Ala Leu ProArg Glu Ser Ser Ser Val Leu Asn Thr Asp Pro Asp 325 330 335 Ala Glu LysPro Arg Phe Ala Ser Asn Leu 340 345 11 1046 DNA Homo sapiens 11cgagtaattt tgtgactgcc atgctgagca agttgagatt tgaggtgttt caacctggag 60attatatcat acgagaagga gccgtgggta aaaaaatgta tttcattcaa cacggtgttg 120ctggtgtcat tacaaaatcc agtaaagaaa tgaagctgac agatggctct tactttggag 180agatttgcct gctgaccaaa ggacgtcgta ctgccagtgt tcgagctgat acatattgtc 240gtctttactc actttccgtg gacaatttca acgaggtcct ggaggaatat ccaatgatga 300ggagagcctt tgagacagtt gccattgacc gactagatcg aataggaaag aaaaattcaa 360ttcttctgca aaagttccag aaggatctga acactggtgt tttcaacaat caggagaacg 420aaatcctcaa gcagattgtg aaacatgaca gggagatggt gcaggcaatc gctcccatca 480attatcctca aatgacaacc ctgaattcca catcgtctac tacgaccccg acctcccgca 540tgaggacaca atctccaccg gtgtacacag cgaccagcct gtctcacagc aacctgcact 600tccccagtcc cagcacacag accccccagc catcagccat cctgtcaccc tgctccacgc 660cgaaaaatga agtgcacaag agcacgcagg cgcttcacaa caccaacctg acccgggaag 720tcaggccact ctccgcctcg cagccctcgc tgccccatga ggtgtccact ctgatttcca 780gacctcatcc cactgtgggc gagtccctgg cctccatccc tcaacccgtg acggcggtcc 840ccggaacggg ccttcaggca gggggcagga gcactgtccc gcagcgcgtc accctcttcc 900gacagatgtc gtcgggagcc atccccccga accgaggagt ccctccagca ccccctccac 960cagcagctgc tcttccaaga gaatcttcct cagtcttaaa cacagaccca gacgcagaaa 1020agccacgatt tgcttcaaat ttatga 1046 12 346 PRT Homo sapiens 12 Asn Phe ValThr Ala Met Leu Ser Lys Leu Arg Phe Glu Val Phe Gln 1 5 10 15 Pro GlyAsp Tyr Ile Ile Arg Glu Gly Ala Val Gly Lys Lys Met Tyr 20 25 30 Phe IleGln His Gly Val Ala Gly Val Ile Thr Lys Ser Ser Lys Glu 35 40 45 Met LysLeu Thr Asp Gly Ser Tyr Phe Gly Glu Ile Cys Leu Leu Thr 50 55 60 Lys GlyArg Arg Thr Ala Ser Val Arg Ala Asp Thr Tyr Cys Arg Leu 65 70 75 80 TyrSer Leu Ser Val Asp Asn Phe Asn Glu Val Leu Glu Glu Tyr Pro 85 90 95 MetMet Arg Arg Ala Phe Glu Thr Val Ala Ile Asp Arg Leu Asp Arg 100 105 110Ile Gly Lys Lys Asn Ser Ile Leu Leu Gln Lys Phe Gln Lys Asp Leu 115 120125 Asn Thr Gly Val Phe Asn Asn Gln Glu Asn Glu Ile Leu Lys Gln Ile 130135 140 Val Lys His Asp Arg Glu Met Val Gln Ala Ile Ala Pro Ile Asn Tyr145 150 155 160 Pro Gln Met Thr Thr Leu Asn Ser Thr Ser Ser Thr Thr ThrPro Thr 165 170 175 Ser Arg Met Arg Thr Gln Ser Pro Pro Val Tyr Thr AlaThr Ser Leu 180 185 190 Ser His Ser Asn Leu His Phe Pro Ser Pro Ser ThrGln Thr Pro Gln 195 200 205 Pro Ser Ala Ile Leu Ser Pro Cys Ser Thr ProLys Asn Glu Val His 210 215 220 Lys Ser Thr Gln Ala Leu His Asn Thr AsnLeu Thr Arg Glu Val Arg 225 230 235 240 Pro Leu Ser Ala Ser Gln Pro SerLeu Pro His Glu Val Ser Thr Leu 245 250 255 Ile Ser Arg Pro His Pro ThrVal Gly Glu Ser Leu Ala Ser Ile Pro 260 265 270 Gln Pro Val Thr Ala ValPro Gly Thr Gly Leu Gln Ala Gly Gly Arg 275 280 285 Ser Thr Val Pro GlnArg Val Thr Leu Phe Arg Gln Met Ser Ser Gly 290 295 300 Ala Ile Pro ProAsn Arg Gly Val Pro Pro Ala Pro Pro Pro Pro Ala 305 310 315 320 Ala AlaLeu Pro Arg Glu Ser Ser Ser Val Leu Asn Thr Asp Pro Asp 325 330 335 AlaGlu Lys Pro Arg Phe Ala Ser Asn Leu 340 345 13 153 DNA Homo sapiens 13ggcggcggcg gcgaggagcc ggcggggggc ttcgaagacg ccgaggggcc ccggcggcag 60tacggcttca tgcagaggca gttcacctcc atgctgcagc ccggggtcaa caaattctcc 120ctccgcatgt ttgggagcca gaaggcggtg gaa 153 14 452 DNA Homo sapiens 14ccgtcgccgg ccgcgtcctc cgggcatgga aggaggcggc aagcccaact cttcgtctaa 60cagccgggac gatggcaaca gcgtcttccc cgccaaggcg tccgcgccgg gcgcggggcc 120ggccgcggcc gagaagcgcc tgggcacccc gccggggggc ggcggggccg gcgcgaagga 180gcacggcaac tccgtgtgct tcaaggtgga cggcggtggc ggcggtggcg gcggcggcgg 240cggcggcgag gagccggcgg ggggcttcga agacgccgag gggccccggc ggcagtacgg 300cttcatgcag aggcagttca cctccatgct gcagcccggg gtcaacaaat tctccctccg 360catgtttggg agccagaagg cggtggaaaa ggagcaggaa agggttaaaa ctgcaggctt 420ctggattatc cacccttaca gtgatttcag gt 452 15 452 DNA Homo sapiens 15ccgtcgccgg ccgcgtcctc cgggcatgga aggaggcggc aagcccaact cttcgtctaa 60cagccgggac gatggcaaca gcgtcttccc cgccaaggcg tccgcgacgg gcgcggggcc 120ggccgcggcc gagaagcgcc tgggcacccc gccggggggc ggcggggccg gcgcgaagga 180gcacggcaac tccgtgtgct tcaaggtgga cggcggtggc ggcggtggcg gcggcggcgg 240cggcggcgag gagccggcgg ggggcttcga agacgccgag gggccccggc ggcagtacgg 300cttcatgcag aggcagttca cctccatgct gcagcccggg gtcaacaaat tctccctccg 360catgtttggg agccagaagg cggtggaaaa ggagcaggaa agggttaaaa ctgcaggctt 420ctggattatc cacccttaca gtgatttcag gt 452 16 142 PRT Homo sapiens 16 MetGlu Gly Gly Gly Lys Pro Asn Ser Ser Ser Asn Ser Arg Asp Asp 1 5 10 15Gly Asn Ser Val Phe Pro Ala Lys Ala Ser Ala Pro Gly Ala Gly Pro 20 25 30Ala Ala Ala Glu Lys Arg Leu Gly Thr Pro Pro Gly Gly Gly Gly Ala 35 40 45Gly Ala Lys Glu His Gly Asn Ser Val Cys Phe Lys Val Asp Gly Gly 50 55 60Gly Gly Gly Gly Gly Gly Gly Gly Gly Gly Glu Glu Pro Ala Gly Gly 65 70 7580 Phe Glu Asp Ala Glu Gly Pro Arg Arg Gln Tyr Gly Phe Met Gln Arg 85 9095 Gln Phe Thr Ser Met Leu Gln Pro Gly Val Asn Lys Phe Ser Leu Arg 100105 110 Met Phe Gly Ser Gln Lys Ala Val Glu Lys Glu Gln Glu Arg Val Lys115 120 125 Thr Ala Gly Phe Trp Ile Ile His Pro Tyr Ser Asp Phe Arg 130135 140 17 142 PRT Homo sapiens 17 Met Glu Gly Gly Gly Lys Pro Asn SerSer Ser Asn Ser Arg Asp Asp 1 5 10 15 Gly Asn Ser Val Phe Pro Ala LysAla Ser Ala Thr Gly Ala Gly Pro 20 25 30 Ala Ala Ala Glu Lys Arg Leu GlyThr Pro Pro Gly Gly Gly Gly Ala 35 40 45 Gly Ala Lys Glu His Gly Asn SerVal Cys Phe Lys Val Asp Gly Gly 50 55 60 Gly Gly Gly Gly Gly Gly Gly GlyGly Gly Glu Glu Pro Ala Gly Gly 65 70 75 80 Phe Glu Asp Ala Glu Gly ProArg Arg Gln Tyr Gly Phe Met Gln Arg 85 90 95 Gln Phe Thr Ser Met Leu GlnPro Gly Val Asn Lys Phe Ser Leu Arg 100 105 110 Met Phe Gly Ser Gln LysAla Val Glu Lys Glu Gln Glu Arg Val Lys 115 120 125 Thr Ala Gly Phe TrpIle Ile His Pro Tyr Ser Asp Phe Arg 130 135 140 18 614 DNA Homo sapiens18 aaccctgaat tccacatcgt ctactacgac cccgacctcc cgcatgagga cacaatctcc 60accggtgtac acagcgacca gcctgtctca cagcaacctg cactccccca gtcccagcac 120acagaccccc cagccatcag ccatcctgtc accctgctcc tacaccaccg cggtctgcag 180ccctcctgta cagagccctc tggccgctcg aactttccac tatgcctccc ccaccgcctc 240ccagctgtca ctcatgcaac agcagccgca gcagcaggta cagcagtccc agccgccgca 300gactcagcca cagcagccgt ccccgcagcc acagacacct ggcagctcca cgccgaaaaa 360tgaagtgcac aagagcacgc aggcgcttca caacaccaac ctgacccggg aagtcaggcc 420actctccgcc tcgcagccct cgctgcccca tgaggtgtcc actctgattt ccagacctca 480tcccactgtg ggcgagtccc tggcctccat ccctcaaccc gtgacggcgg tccccggaac 540gggccttcag gcagggggca ggagcactgt cccgcagcgc gtcaccctct tccgacagat 600gtcgtcggga gcca 614 19 1873 DNA Homo sapiens 19 tgctgcagcc cggggtcaacaaattctccc tccgcatgtt tgggagccag aaggcggtgg 60 aaaaggagca ggaaagggttaaaactgcag gcttctggat tatccaccct tacagtgatt 120 tcaggtttta ctgggatttaataatgctta taatgatggt tggaaatcta gtcatcatac 180 cagttggaat cacattctttacagagcaaa caacaacacc atggattatt ttcaatgtgg 240 catcagatac agttttcctattggacctga tcatgaattt taggactggg actgtcaatg 300 aagacagttc tgaaatcatcctggacccca aagtgatcaa gatgaattat ttaaaaagct 360 ggtttgtggt tgacttcatctcatccatcc cagtggatta tatctttctt attgtagaaa 420 aaggaatgga ttctgaagtttacaagacag ccagggcact tcgcattgtg aggtttacaa 480 aaattctcag tctcttgcgtttattacgac tttcaaggtt aattagatac atacatcaat 540 gggaagagat attccacatgacatatgatc tcgccagtgc agtggtgaga atttttaatc 600 tcatcggcat gatgctgctcctgtgccact gggatggttg tcttcagttc ttagtaccac 660 tactgcagga cttcccaccagattgctggg tgtctttaaa tgaaatggtt aatgattctt 720 ggggaaagca gtattcatacgcactcttca aagctatgag tcacatgctg tgcattgggt 780 atggagccca agccccagtcagcatgtctg acctctggat taccatgctg agcatgatcg 840 tcggggccac ctgctatgccatgtttgtcg gccatgccac cgctttaatc cagtctctgg 900 attcttcgag gcggcagtatcaagagaagt ataagcaagt ggaacaatac atgtcattcc 960 ataagttacc agctgatatgcgtcagaaga tacatgatta ctatgaacac agataccaag 1020 gcaaaatctt tgatgaggaaaatattctca atgaactcaa tgatcctctg agagaggaga 1080 tagtcaactt caactgtcggaaactggtgg ctacaatgcc tttatttgct aatgcggatc 1140 ctaattttgt gactgccatgctgagcaagt tgagatttga ggtgtttcaa cctggagatt 1200 atatcatacg agaaggagccgtgggtaaaa aaatgtattt cattcaacac ggtgttgctg 1260 gtgtcattac aaaatccagtaaagaaatga agctgacaga tggctcttac tttggagaga 1320 tttgcctgct gaccaaaggacgtcgtactg ccagtgttcg agctgataca tattgtcgtc 1380 tttactcact ttccgtggacaatttcaacg aggtcctgga ggaatatcca atgatgagga 1440 gagcctttga gacagttgccattgaccgac tagatcgaat aggaaagaaa aattcaattc 1500 ttctgcaaaa gttccagaaggatctgaaca ctggtgtttt caacaatcag gagaacgaaa 1560 tcctcaagca gattgtgaaacatgacaggg agatggtgca ggcaatcgct cccatcaatt 1620 atcctcaaat gacaaccctgaattccacat cgtctactac gaccccgacc tcccgcatga 1680 ggacacaatc tccaccggtgtacacagcga ccagcctgtc tcacagcaac ctgcactccc 1740 ccagtcccag cacacagaccccccagccat cagccatcct gtcaccctgc tcctacacca 1800 ccgcggtctg cagccctcctgtacagagcc ctctggccgc tcgaactttc cactatgcct 1860 cccccaccgc ctc 1873 201873 DNA Homo sapiens 20 tgctgcagcc cggggtcaac aaattctccc tccgcatgtttgggagccag aaggcggtgg 60 aaaaggagca ggaaagggtt aaaactgcag gcttctggattatccaccct tacagtgatt 120 tcaggtttta ctgggattta ataatgctta taatgatggttggaaatcta gtcatcatac 180 cagttggaat cacattcttt acagagcaaa caacaacaccatggattatt ttcaatgtgg 240 catcagatac agttttccta ttggacctga tcatgaattttaggactggg actgtcaatg 300 aagacagttc tgaaatcatc ctggacccca aagtgatcaagatgaattat ttaaaaagct 360 ggtttgtggt tgacttcatc tcatccatcc cagtggattatatctttctt attgtagaaa 420 aaggaatgga ttctgaagtt tacaagacag ccagggcacttcgcattgtg aggtttacaa 480 aaattctcag tctcttgcgt ttattacgac tttcaaggttaattagatac atacatcaat 540 gggaagagat attccacatg acatatgatc tcgccagtgcagtggtgaga atttttaatc 600 tcatcggcat gatgctgctc ctgtgccact gggatggttgtcttcagttc ttagtaccac 660 tactgcagga cttcccacca gattgctggg tgtctttaaatgaaatggtt aatgattctt 720 ggggaaagca gtattcatac gcactcttca aagctatgagtcacatgctg tgcattgggt 780 atggagccca agccccagtc agcatgtctg acctctggattaccatgctg agcatgatcg 840 tcggggccac ctgctatgcc atgtttgtcg gccatgccaccgctttaatc cagtctctgg 900 attcttcgag gcggcagtat caagagaagt ataagcaagtggaacaatac atgtcattcc 960 ataagttacc agctgatatg cgtcagaaga tacatgattactatgaacac agataccaag 1020 gcaaaatctt tgatgaggaa aatattctca atgaactcaatgatcctctg agagaggaga 1080 tagtcaactt caactgtcgg aaactggtgg ctacaatgcctttatttgct aatgcggatc 1140 ctaattttgt gactgccatg ctgagcaagt tgagatttgaggtgtttcaa cctggagatt 1200 atatcatacg agaaggagcc gtgggtaaaa aaatgtatttcattcaacac ggtgttgctg 1260 gtgtcattac aaaatccagt aaagaaatga agctgacagatggctcttac tttggagaga 1320 tttgcctgct gaccaaagga cgtcgtactg ccagtgttcgagctgataca tattgtcgtc 1380 tttactcact ttccgtggac aatttcaacg aggtcctggaggaatatcca atgatgagga 1440 gagcctttga gacagttgcc attgaccgac tagatcgaataggaaagaaa aattcaattc 1500 ttctgcaaaa gttccagaag gatctgaaca ctggtgttttcaacaatcag gagaacgaaa 1560 tcctcaagca gattgtgaaa catgacaggg agatggtgcaggcaatcgct cccatcaatt 1620 atcctcaaat gacaaccctg aattccacat cgtctactacgaccccgacc tcccgcatga 1680 ggacacaatc tccaccggtg tacacagcga ccagcctgtctcacagcaac ctgcacttcc 1740 ccagtcccag cacacagacc ccccagccat cagccatcctgtcaccctgc tcctacacca 1800 ccgcggtctg cagccctcct gtacagagcc ctctggccgctcgaactttc cactatgcct 1860 cccccaccgc ctc 1873 21 627 DNA Homo sapiens21 cccgggaagt caggccactc tccgcctcgc agccctcgct gccccatgag gtgtccactc 60tgatttccag acctcatccc actgtgggcg agtccctggc ctccatccct caacccgtga 120cggcggtccc cggaacgggc cttcaggcag ggggcaggag cactgtcccg cagcgcgtca 180ccctcttccg acagatgtcg tcgggagcca tccccccgaa ccgaggagtc cctccagcac 240cccctccacc agcagctgct cttccaagag aatcttcctc agtcttaaac acagacccag 300acgcagaaaa gccacgattt gcttcaaatt tatgatccct gctgattgtc aaagcagaaa 360gaaatactct cataaactga gactatactc agatcttatt ttattctatc tcctgataga 420tccctctagc ctactatgaa gagatatttt agacagctgt ggcctacacg tgaaatgtaa 480aaatatatat acatatacta taaaatatat atctaaattc ccaagagagg gtcaaaagac 540ctgtttagca ttcagtgtta tatgtcttcc tttctttaaa tcattaaagg atttaaaatg 600tcaaaaaaaa aaaaaaaggg cggccgc 627 22 2484 DNA Homo sapiens 22 atggaaggaggcggcaagcc caactcttcg tctaacagcc gggacgatgg caacagcgtc 60 ttccccgccaaggcgtccgc gccgggcgcg gggccggccg cggccgagaa gcgcctgggc 120 accccgccggggggcggcgg ggccggcgcg aaggagcacg gcaactccgt gtgcttcaag 180 gtggacggcggtggcggcgg tggcggcggc ggcggcggcg gcgaggagcc ggcggggggc 240 ttcgaagacgccgaggggcc ccggcggcag tacggcttca tgcagaggca gttcacctcc 300 atgctgcagcccggggtcaa caaattctcc ctccgcatgt ttgggagcca gaaggcggtg 360 gaaaaggagcaggaaagggt taaaactgca ggcttctgga ttatccaccc ttacagtgat 420 ttcaggttttactgggattt aataatgctt ataatgatgg ttggaaatct agtcatcata 480 ccagttggaatcacattctt tacagagcaa acaacaacac catggattat tttcaatgtg 540 gcatcagatacagttttcct attggacctg atcatgaatt ttaggactgg gactgtcaat 600 gaagacagttctgaaatcat cctggacccc aaagtgatca agatgaatta tttaaaaagc 660 tggtttgtggttgacttcat ctcatccatc ccagtggatt atatctttct tattgtagaa 720 aaaggaatggattctgaagt ttacaagaca gccagggcac ttcgcattgt gaggtttaca 780 aaaattctcagtctcttgcg tttattacga ctttcaaggt taattagata catacatcaa 840 tgggaagagatattccacat gacatatgat ctcgccagtg cagtggtgag aatttttaat 900 ctcatcggcatgatgctgct cctgtgccac tgggatggtt gtcttcagtt cttagtacca 960 ctactgcaggacttcccacc agattgctgg gtgtctttaa atgaaatggt taatgattct 1020 tggggaaagcagtattcata cgcactcttc aaagctatga gtcacatgct gtgcattggg 1080 tatggagcccaagccccagt cagcatgtct gacctctgga ttaccatgct gagcatgatc 1140 gtcggggccacctgctatgc catgtttgtc ggccatgcca ccgctttaat ccagtctctg 1200 gattcttcgaggcggcagta tcaagagaag tataagcaag tggaacaata catgtcattc 1260 cataagttaccagctgatat gcgtcagaag atacatgatt actatgaaca cagataccaa 1320 ggcaaaatctttgatgagga aaatattctc aatgaactca atgatcctct gagagaggag 1380 atagtcaacttcaactgtcg gaaactggtg gctacaatgc ctttatttgc taatgcggat 1440 cctaattttgtgactgccat gctgagcaag ttgagatttg aggtgtttca acctggagat 1500 tatatcatacgagaaggagc cgtgggtaaa aaaatgtatt tcattcaaca cggtgttgct 1560 ggtgtcattacaaaatccag taaagaaatg aagctgacag atggctctta ctttggagag 1620 atttgcctgctgaccaaagg acgtcgtact gccagtgttc gagctgatac atattgtcgt 1680 ctttactcactttccgtgga caatttcaac gaggtcctgg aggaatatcc aatgatgagg 1740 agagcctttgagacagttgc cattgaccga ctagatcgaa taggaaagaa aaattcaatt 1800 cttctgcaaaagttccagaa ggatctgaac actggtgttt tcaacaatca ggagaacgaa 1860 atcctcaagcagattgtgaa acatgacagg gagatggtgc aggcaatcgc tcccatcaat 1920 tatcctcaaatgacaaccct gaattccaca tcgtctacta cgaccccgac ctcccgcatg 1980 aggacacaatctccaccggt gtacacagcg accagcctgt ctcacagcaa cctgcactcc 2040 cccagtcccagcacacagac cccccagcca tcagccatcc tgtcaccctg ctccacgccg 2100 aaaaatgaagtgcacaagag cacgcaggcg cttcacaaca ccaacctgac ccgggaagtc 2160 aggccactctccgcctcgca gccctcgctg ccccatgagg tgtccactct gatttccaga 2220 cctcatcccactgtgggcga gtccctggcc tccatccctc aacccgtgac ggcggtcccc 2280 ggaacgggccttcaggcagg gggcaggagc actgtcccgc agcgcgtcac cctcttccga 2340 cagatgtcgtcgggagccat ccccccgaac cgaggagtcc ctccagcacc ccctccacca 2400 gcagctgctcttccaagaga atcttcctca gtcttaaaca cagacccaga cgcagaaaag 2460 ccacgatttgcttcaaattt atga 2484 23 2673 DNA Homo sapiens 23 atggaaggag gcggcaagcccaactcttcg tctaacagcc gggacgatgg caacagcgtc 60 ttccccgcca aggcgtccgcgccgggcgcg gggccggccg cggccgagaa gcgcctgggc 120 accccgccgg ggggcggcggggccggcgcg aaggagcacg gcaactccgt gtgcttcaag 180 gtggacggcg gtggcggcggtggcggcggc ggcggcggcg gcgaggagcc ggcggggggc 240 ttcgaagacg ccgaggggccccggcggcag tacggcttca tgcagaggca gttcacctcc 300 atgctgcagc ccggggtcaacaaattctcc ctccgcatgt ttgggagcca gaaggcggtg 360 gaaaaggagc aggaaagggttaaaactgca ggcttctgga ttatccaccc ttacagtgat 420 ttcaggtttt actgggatttaataatgctt ataatgatgg ttggaaatct agtcatcata 480 ccagttggaa tcacattctttacagagcaa acaacaacac catggattat tttcaatgtg 540 gcatcagata cagttttcctattggacctg atcatgaatt ttaggactgg gactgtcaat 600 gaagacagtt ctgaaatcatcctggacccc aaagtgatca agatgaatta tttaaaaagc 660 tggtttgtgg ttgacttcatctcatccatc ccagtggatt atatctttct tattgtagaa 720 aaaggaatgg attctgaagtttacaagaca gccagggcac ttcgcattgt gaggtttaca 780 aaaattctca gtctcttgcgtttattacga ctttcaaggt taattagata catacatcaa 840 tgggaagaga tattccacatgacatatgat ctcgccagtg cagtggtgag aatttttaat 900 ctcatcggca tgatgctgctcctgtgccac tgggatggtt gtcttcagtt cttagtacca 960 ctactgcagg acttcccaccagattgctgg gtgtctttaa atgaaatggt taatgattct 1020 tggggaaagc agtattcatacgcactcttc aaagctatga gtcacatgct gtgcattggg 1080 tatggagccc aagccccagtcagcatgtct gacctctgga ttaccatgct gagcatgatc 1140 gtcggggcca cctgctatgccatgtttgtc ggccatgcca ccgctttaat ccagtctctg 1200 gattcttcga ggcggcagtatcaagagaag tataagcaag tggaacaata catgtcattc 1260 cataagttac cagctgatatgcgtcagaag atacatgatt actatgaaca cagataccaa 1320 ggcaaaatct ttgatgaggaaaatattctc aatgaactca atgatcctct gagagaggag 1380 atagtcaact tcaactgtcggaaactggtg gctacaatgc ctttatttgc taatgcggat 1440 cctaattttg tgactgccatgctgagcaag ttgagatttg aggtgtttca acctggagat 1500 tatatcatac gagaaggagccgtgggtaaa aaaatgtatt tcattcaaca cggtgttgct 1560 ggtgtcatta caaaatccagtaaagaaatg aagctgacag atggctctta ctttggagag 1620 atttgcctgc tgaccaaaggacgtcgtact gccagtgttc gagctgatac atattgtcgt 1680 ctttactcac tttccgtggacaatttcaac gaggtcctgg aggaatatcc aatgatgagg 1740 agagcctttg agacagttgccattgaccga ctagatcgaa taggaaagaa aaattcaatt 1800 cttctgcaaa agttccagaaggatctgaac actggtgttt tcaacaatca ggagaacgaa 1860 atcctcaagc agattgtgaaacatgacagg gagatggtgc aggcaatcgc tcccatcaat 1920 tatcctcaaa tgacaaccctgaattccaca tcgtctacta cgaccccgac ctcccgcatg 1980 aggacacaat ctccaccggtgtacacagcg accagcctgt ctcacagcaa cctgcactcc 2040 cccagtccca gcacacagaccccccagcca tcagccatcc tgtcaccctg ctcctacacc 2100 accgcggtct gcagccctcctgtacagagc cctctggccg ctcgaacttt ccactatgcc 2160 tcccccaccg cctcccagctgtcactcatg caacagcagc cgcagcagca ggtacagcag 2220 tcccagccgc cgcagactcagccacagcag ccgtccccgc agccacagac acctggcagc 2280 tccacgccga aaaatgaagtgcacaagagc acgcaggcgc ttcacaacac caacctgacc 2340 cgggaagtca ggccactctccgcctcgcag ccctcgctgc cccatgaggt gtccactctg 2400 atttccagac ctcatcccactgtgggcgag tccctggcct ccatccctca acccgtgacg 2460 gcggtccccg gaacgggccttcaggcaggg ggcaggagca ctgtcccgca gcgcgtcacc 2520 ctcttccgac agatgtcgtcgggagccatc cccccgaacc gaggagtccc tccagcaccc 2580 cctccaccag cagctgctcttccaagagaa tcttcctcag tcttaaacac agacccagac 2640 gcagaaaagc cacgatttgcttcaaattta tga 2673 24 2484 DNA Homo sapiens 24 atggaaggag gcggcaagcccaactcttcg tctaacagcc gggacgatgg caacagcgtc 60 ttccccgcca aggcgtccgcgacgggcgcg gggccggccg cggccgagaa gcgcctgggc 120 accccgccgg ggggcggcggggccggcgcg aaggagcacg gcaactccgt gtgcttcaag 180 gtggacggcg gtggcggcggtggcggcggc ggcggcggcg gcgaggagcc ggcggggggc 240 ttcgaagacg ccgaggggccccggcggcag tacggcttca tgcagaggca gttcacctcc 300 atgctgcagc ccggggtcaacaaattctcc ctccgcatgt ttgggagcca gaaggcggtg 360 gaaaaggagc aggaaagggttaaaactgca ggcttctgga ttatccaccc ttacagtgat 420 ttcaggtttt actgggatttaataatgctt ataatgatgg ttggaaatct agtcatcata 480 ccagttggaa tcacattctttacagagcaa acaacaacac catggattat tttcaatgtg 540 gcatcagata cagttttcctattggacctg atcatgaatt ttaggactgg gactgtcaat 600 gaagacagtt ctgaaatcatcctggacccc aaagtgatca agatgaatta tttaaaaagc 660 tggtttgtgg ttgacttcatctcatccatc ccagtggatt atatctttct tattgtagaa 720 aaaggaatgg attctgaagtttacaagaca gccagggcac ttcgcattgt gaggtttaca 780 aaaattctca gtctcttgcgtttattacga ctttcaaggt taattagata catacatcaa 840 tgggaagaga tattccacatgacatatgat ctcgccagtg cagtggtgag aatttttaat 900 ctcatcggca tgatgctgctcctgtgccac tgggatggtt gtcttcagtt cttagtacca 960 ctactgcagg acttcccaccagattgctgg gtgtctttaa atgaaatggt taatgattct 1020 tggggaaagc agtattcatacgcactcttc aaagctatga gtcacatgct gtgcattggg 1080 tatggagccc aagccccagtcagcatgtct gacctctgga ttaccatgct gagcatgatc 1140 gtcggggcca cctgctatgccatgtttgtc ggccatgcca ccgctttaat ccagtctctg 1200 gattcttcga ggcggcagtatcaagagaag tataagcaag tggaacaata catgtcattc 1260 cataagttac cagctgatatgcgtcagaag atacatgatt actatgaaca cagataccaa 1320 ggcaaaatct ttgatgaggaaaatattctc aatgaactca atgatcctct gagagaggag 1380 atagtcaact tcaactgtcggaaactggtg gctacaatgc ctttatttgc taatgcggat 1440 cctaattttg tgactgccatgctgagcaag ttgagatttg aggtgtttca acctggagat 1500 tatatcatac gagaaggagccgtgggtaaa aaaatgtatt tcattcaaca cggtgttgct 1560 ggtgtcatta caaaatccagtaaagaaatg aagctgacag atggctctta ctttggagag 1620 atttgcctgc tgaccaaaggacgtcgtact gccagtgttc gagctgatac atattgtcgt 1680 ctttactcac tttccgtggacaatttcaac gaggtcctgg aggaatatcc aatgatgagg 1740 agagcctttg agacagttgccattgaccga ctagatcgaa taggaaagaa aaattcaatt 1800 cttctgcaaa agttccagaaggatctgaac actggtgttt tcaacaatca ggagaacgaa 1860 atcctcaagc agattgtgaaacatgacagg gagatggtgc aggcaatcgc tcccatcaat 1920 tatcctcaaa tgacaaccctgaattccaca tcgtctacta cgaccccgac ctcccgcatg 1980 aggacacaat ctccaccggtgtacacagcg accagcctgt ctcacagcaa cctgcacttc 2040 cccagtccca gcacacagaccccccagcca tcagccatcc tgtcaccctg ctccacgccg 2100 aaaaatgaag tgcacaagagcacgcaggcg cttcacaaca ccaacctgac ccgggaagtc 2160 aggccactct ccgcctcgcagccctcgctg ccccatgagg tgtccactct gatttccaga 2220 cctcatccca ctgtgggcgagtccctggcc tccatccctc aacccgtgac ggcggtcccc 2280 ggaacgggcc ttcaggcagggggcaggagc actgtcccgc agcgcgtcac cctcttccga 2340 cagatgtcgt cgggagccatccccccgaac cgaggagtcc ctccagcacc ccctccacca 2400 gcagctgctc ttccaagagaatcttcctca gtcttaaaca cagacccaga cgcagaaaag 2460 ccacgatttg cttcaaatttatga 2484 25 2673 DNA Homo sapiens 25 atggaaggag gcggcaagcc caactcttcgtctaacagcc gggacgatgg caacagcgtc 60 ttccccgcca aggcgtccgc gacgggcgcggggccggccg cggccgagaa gcgcctgggc 120 accccgccgg ggggcggcgg ggccggcgcgaaggagcacg gcaactccgt gtgcttcaag 180 gtggacggcg gtggcggcgg tggcggcggcggcggcggcg gcgaggagcc ggcggggggc 240 ttcgaagacg ccgaggggcc ccggcggcagtacggcttca tgcagaggca gttcacctcc 300 atgctgcagc ccggggtcaa caaattctccctccgcatgt ttgggagcca gaaggcggtg 360 gaaaaggagc aggaaagggt taaaactgcaggcttctgga ttatccaccc ttacagtgat 420 ttcaggtttt actgggattt aataatgcttataatgatgg ttggaaatct agtcatcata 480 ccagttggaa tcacattctt tacagagcaaacaacaacac catggattat tttcaatgtg 540 gcatcagata cagttttcct attggacctgatcatgaatt ttaggactgg gactgtcaat 600 gaagacagtt ctgaaatcat cctggaccccaaagtgatca agatgaatta tttaaaaagc 660 tggtttgtgg ttgacttcat ctcatccatcccagtggatt atatctttct tattgtagaa 720 aaaggaatgg attctgaagt ttacaagacagccagggcac ttcgcattgt gaggtttaca 780 aaaattctca gtctcttgcg tttattacgactttcaaggt taattagata catacatcaa 840 tgggaagaga tattccacat gacatatgatctcgccagtg cagtggtgag aatttttaat 900 ctcatcggca tgatgctgct cctgtgccactgggatggtt gtcttcagtt cttagtacca 960 ctactgcagg acttcccacc agattgctgggtgtctttaa atgaaatggt taatgattct 1020 tggggaaagc agtattcata cgcactcttcaaagctatga gtcacatgct gtgcattggg 1080 tatggagccc aagccccagt cagcatgtctgacctctgga ttaccatgct gagcatgatc 1140 gtcggggcca cctgctatgc catgtttgtcggccatgcca ccgctttaat ccagtctctg 1200 gattcttcga ggcggcagta tcaagagaagtataagcaag tggaacaata catgtcattc 1260 cataagttac cagctgatat gcgtcagaagatacatgatt actatgaaca cagataccaa 1320 ggcaaaatct ttgatgagga aaatattctcaatgaactca atgatcctct gagagaggag 1380 atagtcaact tcaactgtcg gaaactggtggctacaatgc ctttatttgc taatgcggat 1440 cctaattttg tgactgccat gctgagcaagttgagatttg aggtgtttca acctggagat 1500 tatatcatac gagaaggagc cgtgggtaaaaaaatgtatt tcattcaaca cggtgttgct 1560 ggtgtcatta caaaatccag taaagaaatgaagctgacag atggctctta ctttggagag 1620 atttgcctgc tgaccaaagg acgtcgtactgccagtgttc gagctgatac atattgtcgt 1680 ctttactcac tttccgtgga caatttcaacgaggtcctgg aggaatatcc aatgatgagg 1740 agagcctttg agacagttgc cattgaccgactagatcgaa taggaaagaa aaattcaatt 1800 cttctgcaaa agttccagaa ggatctgaacactggtgttt tcaacaatca ggagaacgaa 1860 atcctcaagc agattgtgaa acatgacagggagatggtgc aggcaatcgc tcccatcaat 1920 tatcctcaaa tgacaaccct gaattccacatcgtctacta cgaccccgac ctcccgcatg 1980 aggacacaat ctccaccggt gtacacagcgaccagcctgt ctcacagcaa cctgcacttc 2040 cccagtccca gcacacagac cccccagccatcagccatcc tgtcaccctg ctcctacacc 2100 accgcggtct gcagccctcc tgtacagagccctctggccg ctcgaacttt ccactatgcc 2160 tcccccaccg cctcccagct gtcactcatgcaacagcagc cgcagcagca ggtacagcag 2220 tcccagccgc cgcagactca gccacagcagccgtccccgc agccacagac acctggcagc 2280 tccacgccga aaaatgaagt gcacaagagcacgcaggcgc ttcacaacac caacctgacc 2340 cgggaagtca ggccactctc cgcctcgcagccctcgctgc cccatgaggt gtccactctg 2400 atttccagac ctcatcccac tgtgggcgagtccctggcct ccatccctca acccgtgacg 2460 gcggtccccg gaacgggcct tcaggcagggggcaggagca ctgtcccgca gcgcgtcacc 2520 ctcttccgac agatgtcgtc gggagccatccccccgaacc gaggagtccc tccagcaccc 2580 cctccaccag cagctgctct tccaagagaatcttcctcag tcttaaacac agacccagac 2640 gcagaaaagc cacgatttgc ttcaaatttatga 2673

1. DNA sequence encoding a full length human hyperpolarised activatedion channel of the HCN 1 subtype or functional equivalents thereof. 2.DNA sequence comprising SEQ ID NO: 22 or SEQ ID NO:
 24. 3. DNA sequenceaccording to SEQ ID NO: 1 or SEQ ID NO:
 5. 4. DNA sequence comprisingSEQ ID NO: 23 or SEQ ID NO:
 25. 5. DNA sequence according to SEQ ID NO:3 or SEQ ID NO:
 7. 6. A full length human hyperpolarised activated ionchannel of the HCN 1 subtype or functional equivalents thereof.
 7. Afull length human hyperpolarised activated ion channel according toclaim 6 encoded by the DNA sequence of claim 2 or
 4. 8. A polypeptidehaving an amino acid sequence comprising SEQ ID NO: 2, SEQ ID NO: 4, SEQID NO: 6 or SEQ ID NO:
 8. 9. Expression vector comprising SEQ ID NO: 22,SEQ ID NO: 24, SEQ ID NO: 23, SEQ ID NO: 25 or fragments thereof. 10.Antibodies reactive with a full length human hyperpolarised activatedion channel according to claims 6 or
 7. 11. Antibodies reactive with apolypeptide according to claim
 8. 12. In vitro screening assay for theselection of compounds that bind and/or influence a function of thehuman hyperpolarised activated ion channel of the HCN 1 subtype. 13.Full length human hyperpolarised activated ion channel of the HCN 1subtype for use in the therapy of disorders selected from the group ofhuman psychiatric and neurological dysfunction of the CNS,cardiovascular dysfunction of the heart, and reproductive dysfunctionand/or contraception related to ¹h function in testes and spermatozoa.14. Compounds selected with an assay according to claim 12 useful in thetreatment of CNS disorders, cardiovascular dysfunction of the heart, andreproductive dysfunction and/or contraception related to Ih function intestes and spermatozoa.
 15. Use of compounds according to claim 14 forthe manufacture of a medicament useful in the treatment of CNSdisorders, cardiovascular dysfunction of the heart, and reproductivedysfunction and/or contraception related to I_(h) function in testes andspermatozoa.